Name: 14441-90-8|5-Phenylisoxazole-3-carboxylic acid|97%
Brand: Ambeed
SKU: A114988
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1
[ 14441-90-8 ]
[ 5071-61-4 ]

Reference： [1]Gazzetta Chimica Italiana,1940,vol. 70,p. 240,244

2
[ 7063-99-2 ]
[ 14441-90-8 ]

Yield	Reaction Conditions	Operation in experiment
94%		Synthesis of 5-phenyl-isoxazole-3-carboxylic acid (57) A solution of phenyl-substituted isoxazole ethyl ester (53) (1.89 g, 8.70 mmol) in ethanol (30 mL) was stirred at room temperature. To this solution was added a 2M NaOH solution (6.5 mL, 13.1 mmol). After 5 min. TLC showed that the reaction was complete. To the reaction mixture was added 0.5M HCl to adjust the pH to 3-4, before extracting with ethyl acetate (2×75 mL). The organic extracts were combined, washed with brine, dried over sodium sulfate, and concentrated to afford 5-phenyl-isoxazole-3-carboxylic acid (57) was obtained as a white solid (1.54 g, 94%). 57: 1H NMR (500 MHz, CDCl3): delta 9.4 (broad, 1H), 7.83 (d, 2H), 7.51 (m, 3H), 6.99 (s, 1H)
94%		A solution of phenyl-substituted isoxazole ethyl ester (53) (1.89 g, 8.70 mmol) in ethanol (30 mL) was stirred at room temperature. To this solution was added a 2 M NaOH solution (6.5 mL, 13.1 mmol). After 5 min, TLC showed that the reaction was complete. To the reaction mixture was added 0.5 M HCl to adjust the pH to 34, before extracting with ethyl acetate (2×75 mL). The organic extracts were combined, washed with brine, dried over sodium sulfate, and concentrated to afford 5-phenyl-isoxazole-3-carboxylic acid (57) obtained as a white solid (1.54 g, 94%). 57: 1H NMR (500 MHz, CDCl3): delta 9.4 (broad, 1H), 7.83 (d, 2H), 7.51 (m, 3H), 6.99 (s, 1H)
91.5%	With sodium hydroxide; In methanol; at 20℃; for 4h;	General procedure: Sodium hydroxide (2N) was added to a solution of intermediate 2a-i (1 equiv.) in methanol at ambient temperature. The reaction mixture was stirred for 4h and the methanol was removed by rotary evaporation. The resultant mixture was adjusted to pH=5-6 with 1N HCl solution. The precipitated white solid was collected by filtration and dried to give the carboxylic acid intermediate (1a-i). 4.13.1 5-Phenylisoxazole-3-carboxylic acid(1a) (0032) Light white solid; yield: 91.5%; 1H NMR (600MHz, DMSO-d6) delta 7.95 (dd, J=7.8, 1.7Hz, 2H), 7.58-7.53 (m, 3H), 7.41 (s, 1H).

79%		General procedure: To a stirred solution of ester 12-20 (1 equiv) in EtOH (95%), was added sodium hydroxide in pellets (10 equiv). The mixture was then stirred at room temperature for 24 h. EtOH was removed under reduced pressure and the residue was acidified (1N HCl, pH 2) and extracted with EtOAc. The organic extracts were washed with water and brine, dried over MgSO4 and concentrated under reduced pressure to afford essentially pure carboxylic acid 21-29.
79%	With sodium hydroxide; In ethanol; at 20℃; for 24h;	General procedure: To a stirred solution of ester 22-32 (1 equiv) in 95% EtOH (50 mL), was added sodium hydroxide in pellets (10 equiv). The mixture was stirred at room temperature for 24 h. EtOH was removed under reduced pressure and the residue was acidified (1 N HCl, pH 2) and extracted with EtOAc (220 mL). The organic layers were washed with water (20 mL) and brine (20 mL), dried over MgSO4 and evaporated under reduced pressure to afford pure carboxylic acids 33-43.
73%	With lithium hydroxide monohydrate; In tetrahydrofuran; methanol; water; at 20℃; for 4h;	General procedure: To a solution of the obtained ethyl ester intermediate (1 equiv) in 2:3:1 THF/MeOH/H2O (18 ml) was added LiOH·H2O (1.5 equiv). After stirring at room temperature for 4 h, the volatiles were removed under reduced pressure. The residue was acidified with 1N hydrochloric acid solution, and then filtered and the filter cake was washed with 5 mL of water, dried in vacuum to afford a white powder. Recrystallization from 75% EtOH gave the desired compounds 1-8.
69%	With lithium hydroxide monohydrate; water; In tetrahydrofuran; at 20℃; for 2h;	Acetophenone (3 g, 25 mmol) was taken up in 30 mL of dry toluene and NaH (780 mg, 32 mmol) was then added. The resulting reaction mixture was stirred at room temperaturefor 60 minutes. A solution of diethyl oxalate (5.5 g, 37.5 mmol) in dry toluene (25 mL) was then added drop wise and stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and the resulting residue was diluted with ice water. The precipitated solids were collected by filtration and dried to afford 2.85 g of ethyl 2,4-dioxo-4-phenylbutanoate (52%yield) as a yellow solid. This material (2.85 g, 12.9 mmol) was taken up in EtOH (25 mL) along with NH2OH.HC1 (1.16 g, 16.8 mmol) and then stirred under reflux for 3 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was diluted with water and extracted with EtOAc. The combined organic layers were washed with H20, dried (Na2SO4) and concentrated under reduced pressure. Purification by silica gelchromatography (pentanes/EtOAc) afforded ethyl 5-phenylisoxazole-3 -carboxylate (2.53 g,90% yield) as a white solid. This material (2.53 g, 11.6 mmol) was taken up in THF/H20 (45 mL/5 mL) along with LiOH.H20 (1.0 g, 23.3 mmol) and the resulting reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then concentrated under reduced pressure. Sufficient 1 N HC1 was added to the resulting residue to bring the pH toabout 5. The resulting solids were collected by filtration and dried under high vacuum to afford1.5 g of 5-phenylisoxazole-3-carboxylic acid (69%) as a white solid. 5-Phenylisoxazole-3- carboxylic acid was then coupled with (4Z,7Z,1OZ,13Z,16Z,19Z)-N-(2-(((R)-3-amino-4-((1,3- dihydroxypropan-2-yl)amino)-2-methyl-4-oxobutan-2-yl)disulfanyl)ethyl)docosa- 4,7,10,13,16,19-hexaenamide using the same general amide coupling procedure describedearlier (see example 8) to obtain N-((R)-1-((1,3-dihydroxypropan-2-yl)amino)-3-((2- ((4Z,7Z, 1 OZ, 1 3Z, 1 6Z, 1 9Z)-docosa-4,7, 10,13,16,1 9-hexaenamido)ethyl)disulfanyl)-3 -methyl-ioxobutan-2-yl)-5-phenylisoxazole-3 -carboxamide. MS (El) calc? d for C49H58N40652 778.38; found 779 [M+H].
69%	With lithium hydroxide monohydrate; water; In tetrahydrofuran; at 20℃; for 2h;	10298] Acetophenone (3 g, 25 mmol) was taken up in 30 mE of dry toluene and NaR (780 mg, 32 mmol) was then added. The resulting reaction mixture was stirred at room temperature for 60 minutes. A solution of diethyl oxalate (5.5 g, 37.5 mmol) in dry toluene (25 mE) was then added drop wise and stirred at room temperature for 1 hout The reaction mixture was concentrated under reduced pressure and the resulting residue was diluted with ice watet The precipitated solids were collected by filtration and dried to afford 2.85 g of ethyl 2,4-dioxo-4-phenylbutanoate (52% yield) as a yellow solid. This material (2.85 g, 12.9 mmol) was taken up in EtOR (25 mE) along with NH2OH.HC1 (1.16 g, 16.8 mmol) and then stirred under reflux for 3 hours. The reaction mixture was concentrated under reduced presresulting sure. The resulting residue was diluted with water and extracted with EtOAc. The combined organic layers were washed with H20, dried (Na2SO4) and concentrated under reduced pressure. Purification by silica gel chromatography (pentanes/EtOAc) afforded ethyl 5-phenylisoxazole-3-car- boxylate (2.53 g, 90% yield) as a white solid. This material (2.53 g, 11.6 mmol) was taken up in THF/H20 (45 mE/S mE) along with EiOH.H20 (1.0 g, 23.3 mmol) and thereaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then concentrated under reduced pressure. Sufficient 1 N HC1 was added to the resulting residue to bring the pH to about 5. The resulting solids were collected by filtration and dried under high vacuum to afford 1.5 g of 5-phenylisoxazole-3-carboxylic acid (69%) as a white solid. 5-Phenylisoxazole-3-carboxylic acid was then coupled with (4Z,7Z,10Z,13Z,16Z,19Z)- N-((R)-1 -amino-3-(((R)-3-amino-4-((1 ,3-dihydroxypro- pan-2-yl)amino)-2-methyl-4-oxobutan-2-yl)disulfanyl)-1 - oxopropan-2-yl)docosa-4,7, 10,13,16,1 9-hexaenamide using the same amide coupling procedure as detailed in example 4. The final product was purified by silica gel chromatography (CH2C12/MeOH). MS, calculated for C43H59N50752:821.39; found 822 [M+H].
24 g	With lithium hydroxide; In tetrahydrofuran; water; for 2h;	To a stirred solution of ethyl 5-phenylisoxazole-3- carboxylate (28.0 g, 129mmol) in THF (200mL) was added lithium hydroxide (21.16g, 516mmol) in water (200mL). The reaction was stirred for 2h. The organic solvent was distilled off, water was added (500 mL), and acidified with aq. 5N HC1 (50mL). The solid precipitate was collected by filtration and dried under vacuum to give 5-phenylisoxazole-3- carboxylic acid (24.0 g, 190 [M+H]); 'H NMR: (400 MHz, DMSO) delta: 7.438(S, 1H), 7.524- 7.593(m,3H), 7.945-7.969(m, 2H), 14.1 15(S, 1H).

Reference： [1]Patent: US2006/199853,2006,A1 .Location in patent: Page/Page column 45
[2]Patent: US2006/223884,2006,A1 .Location in patent: Page/Page column 58
[3]ACS Medicinal Chemistry Letters,2020,vol. 11,p. 605 - 610
[4]European Journal of Medicinal Chemistry,2017,vol. 137,p. 96 - 107
[5]Bioorganic and Medicinal Chemistry,2011,vol. 19,p. 3777 - 3786
[6]Bioorganic and Medicinal Chemistry,2013,vol. 21,p. 5383 - 5394
[7]Bioorganic and Medicinal Chemistry Letters,2019,vol. 29,p. 525 - 528
[8]Patent: WO2016/86136,2016,A1 .Location in patent: Paragraph 00226
[9]Patent: US2017/342046,2017,A1 .Location in patent: Paragraph 0297; 0298
[10]Gazzetta Chimica Italiana,1948,vol. 78,p. 630,636
[11]Gazzetta Chimica Italiana,1938,vol. 68,p. 566,568
[12]Journal of the Chemical Society. Perkin transactions I,1972,p. 437 - 443
[13]Journal of Medicinal Chemistry,2008,vol. 51,p. 4370 - 4373
[14]Letters in Organic Chemistry,2010,vol. 7,p. 32 - 38
[15]Journal of Heterocyclic Chemistry,2015,vol. 52,p. 1743 - 1747
[16]Patent: WO2016/54560,2016,A1 .Location in patent: Page/Page column 36
[17]Letters in drug design and discovery,2017,vol. 14,p. 58 - 65
[18]Chemistry and biodiversity,2019,vol. 16

3
[ 25741-97-3 ]
[ 124-38-9 ]
[ 14441-90-8 ]

Reference： [1]Gazzetta Chimica Italiana,1969,vol. 99,p. 1107 - 1114

4
[ 14441-90-8 ]
[ 5932-30-9 ]

Reference： [1]Gazzetta Chimica Italiana,1940,vol. 70,p. 240,244

5
[ 892840-58-3 ]
[ 14441-90-8 ]
5-phenyl-isoxazole-3-carboxylic acid [2-(3-dimethylamino-pyrrolidin-1-yl)-benzothiazol-6-yl]-amide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
71%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 40℃;	6 Combine 2-(3-dimethylamino-pyrrolidin-l-yl)-benzothiazol-6-ylamine (0.10Og, 0.381 mmol) (Example 1, step 3), dissolved in CH2Cl2 (5.0 mL), with S-phenyl-isoxazole-S-carboxylic acid (0.079 g, 0.419 mmol), HATU (0.145 g, 0.381 mmol), and DIEA (0.20 mL, 1.14 mmol) in a 40 mL reaction vial and shake the mixture overnight at 40 0C. Dilute the mixture with CH2Cl2 (25 mL) and wash with 1.0 M NaOH (25 mL) which results in emulsions forming. Wait 3 h for the emulsions to disappear, separate the layers and dry the organic portion over sodium sulfate. Filter and concentrate in vacuo to yield the compound as a yellow solid. Purify using silica gel chromatography (5% MeOH/CH2Cl2) to afford the title compound as a yellowish-white solid (117 mg, 71%). mass spectrum (m/e): 434.2 [M+l], 432.2 [M-I]. 1H NMR (400 MHz, DMSO- d6): δ 10.76 (s, IH), 8.27 (d, IH, J = 2 Hz), 7.98 (m, 2H), 7.56-7.63 (m, 4H), 7.49 (s, IH), 7.45 (d, IH, / = 8.8 Hz), 3.73 (t, IH, J = 9.2 Hz), 3.64 (t, IH, J = 9.2 Hz), 3.49 (m, IH), 3.29 (m, IH), 2.90 (m, IH), 2.21 (s, 6H), 2.19 (m, IH), 1.91 (m, IH).

Reference： [1]Current Patent Assignee: ELI LILLY & CO - WO2006/66173, 2006, A2 Location in patent: Page/Page column 36

6
[ 14441-90-8 ]
[ 1619-37-0 ]

Yield	Reaction Conditions	Operation in experiment
96%	With lithium aluminium tetrahydride In tetrahydrofuran at 0 - 20℃; for 0.5h;	73 To a stirred suspension of LAH in THF (10 mL, 1M solution in THF) at 0° C. was added a solution of 5-phenylisoxazole-3-carboxylic acid (0.9 g in 5 mL of THF) and after completion of addition the reaction was slowly warmed to room temperature (30 min). The reaction mixture was cooled to 0° C. and ethyl acetate was added (30 mL), followed by slow addition of saturated sodium sulfate solution. The solid was rinsed with ether several times and the solvent decanted. The combined organic layer was dried over MgSO4, filtered and concentrated in vacuo to get (5-phenylisoxazol-3-yl)methanol (0.8 g, 96% yield).
	With borane-THF In tetrahydrofuran at 80℃; for 10h;	4.1 1) Production of (5-phenylisoxazol-3-yl)methanol To a solution of 5-phenylisoxazol-3-carboxylic acid (551 mg) in tetrahydrofuran (15 ml), 3.7 ml of borane-tetrahydrofuran complex (1.17M tetrahydrofuran solution) was added, and the reaction solution was stirred at 80°C for 10 hours. Water and a saturated aqueous sodium hydrogen carbonate solution were added to the reaction solution, and the mixture was extracted with chloroform and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue obtained was purified by silica gel column chromatography (eluent: ethyl acetate/hexane (10:90-50:50)) to afford the title compound as a white solid.

Reference： [1]Current Patent Assignee: ABBVIE INC - US2006/211603, 2006, A1 Location in patent: Page/Page column 65
[2]Current Patent Assignee: MERCK & CO INC - EP2098517, 2009, A1 Location in patent: Page/Page column 29

7
[ 666859-55-8 ]
[ 14441-90-8 ]
[ 666859-30-9 ]

Yield	Reaction Conditions	Operation in experiment
20%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With oxalyl dichloride In DMF (N,N-dimethyl-formamide); dichloromethane at 20℃; for 6h; Stage #2: 3-(2-amino-5-bromophenyl)-4,5-dihydro-1,2,4-oxadiazol-5-one In pyridine; dichloromethane	30 N-[4-bromo-2-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)phenyl-5-[phenylisoxazole-3-carboxamide PHA-689663 5-phenylisoxazole-3-carboxylic acid [(SHIMIZU,] T.; Hayashi, Y.; Teramura, K. Bull.[CHEM..] Soc. [JPN.] 1985, 58, 2519; 245 mg, 1.29 mmol), DCM (40 ml) and oxyl chloride (4.0 ml) were place in a flask, followed by the addition of on drop of DMF (0.1 ml). The solution was stirred at room temperature for about 6 h. Then heptane (20 ml) was added and the solvent was removed. The residue was dissolved in DCM (10 ml). [3- (2-amino-5-bromophenyl)-1,] 2,4-oxadiazol-5 [(4H)-ONE] (300 mg, [1.] [18] mmol) was added and followed by the addition of pyridine (0.3 ml). The resulting solution was stirred overnight, then diluted with MTBE (200 ml) and washed with IN [HC1,] brine, dried [(MGS04),] filtered, and concentrated in vacuo. The residue was purified by flash chromatography [(DCM/MEOH=] 1: 0, [50] : 1) to afford 105 mg (20%) of the desired product as a white [SOLID. 1H] NMR (300 MHz, DMSO-d6) 8 11.03 (s, [1] H), 8.27 (d, J= 9.0 Hz, [1] H), 8.00 [(M,] 2 H), 7.97 (d, J= 2.3 Hz, 1 H), 7.89 (dd, J= 2.3, 8. [8 HZ,] 1 H), 7.57 [(M,] 4 H).

Reference： [1]Current Patent Assignee: PFIZER INC - WO2004/18461, 2004, A2 Location in patent: Page 46-47

8
[ 14441-90-8 ]
[ 1086562-05-1 ]
[ 1086562-36-8 ]

Yield	Reaction Conditions	Operation in experiment
94%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With oxalyl dichloride; N,N-dimethyl-formamide In dichloromethane at 20℃; Stage #2: 2-(2-amino-3-methoxyphenyl)-benzoxazole-4-carboxylic acid methyl ester With pyridine In dichloromethane

Reference： [1]Tipparaju, Suresh K.; Joyasawal, Sipak; Pieroni, Marco; Kaiser, Marcel; Brun, Reto; Kozikowski, Alan P. [Journal of Medicinal Chemistry, 2008, vol. 51, # 23, p. 7344 - 7347]

9
[ 14441-90-8 ]
[ 2038-57-5 ]
[ 1314023-88-5 ]

Yield	Reaction Conditions	Operation in experiment
67%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 0.75h; Stage #2: 3-Phenylpropan-1-amine In dichloromethane at 20℃; for 24h;	4.1.5. General procedure for the preparation of 5-aryl-isoxazole-3-carboxamide (30-48) General procedure: To a solution of carboxylic acid 21-29 (1 equiv) in anhydrous CH2Cl2 were successively added HBTU (1.5 equiv), HOBt (0.5 equiv) and DIPEA (2 equiv). The mixture was stirred for 45 min at room temperature. Then, the appropriate amine (1.1 equiv) was introduced and the stirring was continued for 24 h. At the end of the reaction, the mixture was filtered off and the filtrate was successively washed with saturated aqueous NaHCO3 solution, 1N aqueous HCl and distilled water. The organic layer was dried over MgSO4 and was concentrated in vacuo. The resulting residue was purified by TLC (cyclohexane/AcOEt, 7:3) and crystallized in absolute EtOH to give carboxamide 30-48.
	With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

Reference： [1]Location in patent: experimental part Andrzejak, Virginie; Muccioli, Giulio G.; Body-Malapel, Mathilde; El Bakali, Jamal; Djouina, Madjid; Renault, Nicolas; Chavatte, Philippe; Desreumaux, Pierre; Lambert, Didier M.; Millet, Regis [Bioorganic and Medicinal Chemistry, 2011, vol. 19, # 12, p. 3777 - 3786]
[2]Location in patent: body text Andrzejak, Virginie; Millet, Regis; Bakali, Jamal El; Guelzim, Abdelhalim; Gluszok, Sebastien; Chavatte, Philippe; Bonte, Jean-Paul; Vaccher, Claude; Lipka, Emmanuelle [Letters in Organic Chemistry, 2010, vol. 7, # 1, p. 32 - 38]

10
[ 14441-90-8 ]
[ 18107-18-1 ]
[ 51677-09-9 ]

Yield	Reaction Conditions	Operation in experiment
	With methanol; In hexanes; toluene; at 20℃;	Example 9; 1 -(4-(5-(4-( 1 , 1 -Difluoroethyl)-5-phenylisoxazol-3 -yl)- 1 ,2,4-oxadiazol-3 - yl)benzyl)azetidine-3-carboxylic acid, 2,2,2-trifluoroacetic acid salt; 9-A. Methyl S-phenylisoxazole-S-carboxylate; [00179] A solution of S-phenylisoxazole-S-carboxylic acid (0.86 g, 4.55 mmol) in toluene (15.0 mL) and methanol (3 mL) was added a 2M solution of TMS- diazomethane in hexanes (3.1 mL, 6.14 mmol) dropwise at room temperature. The reaction mixture was stirred for 30 minutes and concentrated. The residue was dispersed in methanol (3 mL), stirred for 5 minutes, and filtered to give methyl 5- phenylisoxazole-3-carboxylate (897 mg). The compound had an HPLC ret. time = 2.67 min. : YMC S5 COMBISCREEN 4.6X50 mm; Gradient time: 4 min; Flow rate = 4 ml/min; Solvent A = 10% MeOH - 90% Water - 0.2% H3PO4; Solvent B = 90% MeOH - 10% water - 0.2% H3PO4; Start % B = O; Final % B = 100. LC-MS: M+1 = 204+.

Reference： [1]Patent: WO2010/85581,2010,A1 .Location in patent: Page/Page column 82
[2]Journal of Medicinal Chemistry,2016,vol. 59,p. 2820 - 2840

11
[ 14441-90-8 ]
[ 1484-85-1 ]
[ 1314023-95-4 ]

Yield	Reaction Conditions	Operation in experiment
65%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 0.75h; Stage #2: 3,4-methylenedioxyphenylethylamine In dichloromethane at 20℃; for 24h;	4.1.5. General procedure for the preparation of 5-aryl-isoxazole-3-carboxamide (30-48) General procedure: To a solution of carboxylic acid 21-29 (1 equiv) in anhydrous CH2Cl2 were successively added HBTU (1.5 equiv), HOBt (0.5 equiv) and DIPEA (2 equiv). The mixture was stirred for 45 min at room temperature. Then, the appropriate amine (1.1 equiv) was introduced and the stirring was continued for 24 h. At the end of the reaction, the mixture was filtered off and the filtrate was successively washed with saturated aqueous NaHCO3 solution, 1N aqueous HCl and distilled water. The organic layer was dried over MgSO4 and was concentrated in vacuo. The resulting residue was purified by TLC (cyclohexane/AcOEt, 7:3) and crystallized in absolute EtOH to give carboxamide 30-48.

Reference： [1]Location in patent: experimental part Andrzejak, Virginie; Muccioli, Giulio G.; Body-Malapel, Mathilde; El Bakali, Jamal; Djouina, Madjid; Renault, Nicolas; Chavatte, Philippe; Desreumaux, Pierre; Lambert, Didier M.; Millet, Regis [Bioorganic and Medicinal Chemistry, 2011, vol. 19, # 12, p. 3777 - 3786]

12
[ 14441-90-8 ]
[ 143-16-8 ]
[ 1334687-01-2 ]

Yield	Reaction Conditions	Operation in experiment
50%		General procedure: Thionyl chloride (1.2 mL) was added at 0 C to the 3-phenylisoxazole-5-carboxylic acid 13 or the 5-phenylisoxazole-3-carboxylic acid 14 or the 5-(4-chlorophenyl)isoxazole-3-carboxylic acid 15 or the 3-(4-methylphenyl)isoxazole-5-carboxylic acid 16 (0.3 g, 1.58 mmol). The obtained suspension was stirred and heated at reflux for 16 h and then cooled at 0 C. At this temperature, a new addition of thionyl chloride (1.2 mL) was followed by another heating at reflux for 2 h. The reaction mixture was cooled at room temperature and the thionyl chloride excess was evaporated to dryness in vacuo. Anhydrous THF (2 mL) was added to the crude product. To the resulting solution, cooled at -5 C, was added dropwise a solution of appropriate amine (3.16 mmol) in dry THF (2 mL). The reaction mixture was stirred at room temperature for ca. 90 min (TLC, petroleum ether/ethyl acetate) and then the solid mass was filtered off and washed with THF. The filtrate was evaporated to dryness; the residue was treated with saturated sodium bicarbonate solution (20 mL) and extracted with dichloromethane (3×15 mL). The combined organic phases were dried (Na2SO4) and evaporated to dryness to give the crude product, purified by flash-chromatography to give the desired amide.

Reference： [1]European Journal of Medicinal Chemistry,2011,vol. 46,p. 4506 - 4520

13
[ 14441-90-8 ]
[ 142-84-7 ]
[ 1328832-53-6 ]

Yield	Reaction Conditions	Operation in experiment
25%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With thionyl chloride at 0℃; Reflux; Stage #2: di-n-propylamine In tetrahydrofuran at -5 - 20℃;	6.2.3. General procedure for the synthesis of N,N-dialkyl-3-arylisoxazole-5-carboxamides 3b-e,s and N,N-dialkyl-5-arylisoxazole-3-carboxamides 4b-e,h,k General procedure: Thionyl chloride (1.2 mL) was added at 0 °C to the 3-phenylisoxazole-5-carboxylic acid 13 or the 5-phenylisoxazole-3-carboxylic acid 14 or the 5-(4-chlorophenyl)isoxazole-3-carboxylic acid 15 or the 3-(4-methylphenyl)isoxazole-5-carboxylic acid 16 (0.3 g, 1.58 mmol). The obtained suspension was stirred and heated at reflux for 16 h and then cooled at 0 °C. At this temperature, a new addition of thionyl chloride (1.2 mL) was followed by another heating at reflux for 2 h. The reaction mixture was cooled at room temperature and the thionyl chloride excess was evaporated to dryness in vacuo. Anhydrous THF (2 mL) was added to the crude product. To the resulting solution, cooled at -5 °C, was added dropwise a solution of appropriate amine (3.16 mmol) in dry THF (2 mL). The reaction mixture was stirred at room temperature for ca. 90 min (TLC, petroleum ether/ethyl acetate) and then the solid mass was filtered off and washed with THF. The filtrate was evaporated to dryness; the residue was treated with saturated sodium bicarbonate solution (20 mL) and extracted with dichloromethane (3×15 mL). The combined organic phases were dried (Na2SO4) and evaporated to dryness to give the crude product, purified by flash-chromatography to give the desired amide.

Reference： [1]Location in patent: experimental part Cosimelli, Barbara; Simorini, Francesca; Taliani, Sabrina; La Motta, Concettina; Da Settimo, Federico; Severi, Elda; Greco, Giovanni; Novellino, Ettore; Costa, Barbara; Da Pozzo, Eleonora; Bendinelli, Sara; Martini, Claudia [European Journal of Medicinal Chemistry, 2011, vol. 46, # 9, p. 4506 - 4520]

14
[ 14441-90-8 ]
[ 111-92-2 ]
[ 1334686-99-5 ]

Yield	Reaction Conditions	Operation in experiment
30%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With thionyl chloride at 0℃; Reflux; Stage #2: dibutylamine In tetrahydrofuran at -5 - 20℃;	6.2.3. General procedure for the synthesis of N,N-dialkyl-3-arylisoxazole-5-carboxamides 3b-e,s and N,N-dialkyl-5-arylisoxazole-3-carboxamides 4b-e,h,k General procedure: Thionyl chloride (1.2 mL) was added at 0 °C to the 3-phenylisoxazole-5-carboxylic acid 13 or the 5-phenylisoxazole-3-carboxylic acid 14 or the 5-(4-chlorophenyl)isoxazole-3-carboxylic acid 15 or the 3-(4-methylphenyl)isoxazole-5-carboxylic acid 16 (0.3 g, 1.58 mmol). The obtained suspension was stirred and heated at reflux for 16 h and then cooled at 0 °C. At this temperature, a new addition of thionyl chloride (1.2 mL) was followed by another heating at reflux for 2 h. The reaction mixture was cooled at room temperature and the thionyl chloride excess was evaporated to dryness in vacuo. Anhydrous THF (2 mL) was added to the crude product. To the resulting solution, cooled at -5 °C, was added dropwise a solution of appropriate amine (3.16 mmol) in dry THF (2 mL). The reaction mixture was stirred at room temperature for ca. 90 min (TLC, petroleum ether/ethyl acetate) and then the solid mass was filtered off and washed with THF. The filtrate was evaporated to dryness; the residue was treated with saturated sodium bicarbonate solution (20 mL) and extracted with dichloromethane (3×15 mL). The combined organic phases were dried (Na2SO4) and evaporated to dryness to give the crude product, purified by flash-chromatography to give the desired amide.

Reference： [1]Location in patent: experimental part Cosimelli, Barbara; Simorini, Francesca; Taliani, Sabrina; La Motta, Concettina; Da Settimo, Federico; Severi, Elda; Greco, Giovanni; Novellino, Ettore; Costa, Barbara; Da Pozzo, Eleonora; Bendinelli, Sara; Martini, Claudia [European Journal of Medicinal Chemistry, 2011, vol. 46, # 9, p. 4506 - 4520]

15
[ 14441-90-8 ]
[ 2050-92-2 ]
[ 1334687-00-1 ]

Yield	Reaction Conditions	Operation in experiment
79%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With thionyl chloride at 0℃; Reflux; Stage #2: Di-n-amylamine In tetrahydrofuran at -5 - 20℃;	6.2.3. General procedure for the synthesis of N,N-dialkyl-3-arylisoxazole-5-carboxamides 3b-e,s and N,N-dialkyl-5-arylisoxazole-3-carboxamides 4b-e,h,k General procedure: Thionyl chloride (1.2 mL) was added at 0 °C to the 3-phenylisoxazole-5-carboxylic acid 13 or the 5-phenylisoxazole-3-carboxylic acid 14 or the 5-(4-chlorophenyl)isoxazole-3-carboxylic acid 15 or the 3-(4-methylphenyl)isoxazole-5-carboxylic acid 16 (0.3 g, 1.58 mmol). The obtained suspension was stirred and heated at reflux for 16 h and then cooled at 0 °C. At this temperature, a new addition of thionyl chloride (1.2 mL) was followed by another heating at reflux for 2 h. The reaction mixture was cooled at room temperature and the thionyl chloride excess was evaporated to dryness in vacuo. Anhydrous THF (2 mL) was added to the crude product. To the resulting solution, cooled at -5 °C, was added dropwise a solution of appropriate amine (3.16 mmol) in dry THF (2 mL). The reaction mixture was stirred at room temperature for ca. 90 min (TLC, petroleum ether/ethyl acetate) and then the solid mass was filtered off and washed with THF. The filtrate was evaporated to dryness; the residue was treated with saturated sodium bicarbonate solution (20 mL) and extracted with dichloromethane (3×15 mL). The combined organic phases were dried (Na2SO4) and evaporated to dryness to give the crude product, purified by flash-chromatography to give the desired amide.

Reference： [1]Location in patent: experimental part Cosimelli, Barbara; Simorini, Francesca; Taliani, Sabrina; La Motta, Concettina; Da Settimo, Federico; Severi, Elda; Greco, Giovanni; Novellino, Ettore; Costa, Barbara; Da Pozzo, Eleonora; Bendinelli, Sara; Martini, Claudia [European Journal of Medicinal Chemistry, 2011, vol. 46, # 9, p. 4506 - 4520]

16
[ 14441-90-8 ]
[ 100-51-6 ]
[ 1364667-00-4 ]

Yield	Reaction Conditions	Operation in experiment
27%	With dmap; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In acetonitrile at 20℃; for 12h;	Benzyl 5-phenylisoxazole-3-carboxylate (7) 5-Phenylisoxazole-3-carboxylic acid (purchased from PharamCore, http://www.pharmacore.com; 100 mg, 0.50 mmol) and benzyl alcohol (79 mL, 0.75 mmol) were dissolved in 3 mL of acetonitrile and treated with EDC (144 mg, 0.75 mmol) and DMAP (184 mg, 1.51 mmol). The resulting solution was stirred for 12 h at room temperature. The reaction mixture was diluted with CH2Cl2, washed with 10% aqueous NaHCO3 solution (2x), water and 5% acetic acid solution (2x). The organic phase was collected, dried over Na2SO4, filtered and then concentrated in vacuo. Crude material obtained was recrystallized with hot acetonitrile to give 38 mg (27%) of 7 as a white solid. Mp 95-96oC; 1H NMR (300 MHz, CDCl3) 7.82 - 7.78 (2 H, m), 7.51-7.33 (8 H, m), 6.93 (1 H, s), 5.44 (2 H, s); 13C NMR (126 MHz, CDCl3) d 172.00, 160.06, 156.94, 135.07, 131.03, 129.35, 128.90, 128.88, 128.83, 126.78, 126.13, 100.18, 67.91. HRMS (EI), M+1 calcd. for C17H14NO3, 280.0968; found 280.1008. HPLC tR = 10.5 min.

Reference： [1]Moraski, Garrett C.; Markley, Lowell D.; Chang, Mayland; Cho, Sanghyun; Franzblau, Scott G.; Hwang, Chang Hwa; Boshoff, Helena; Miller, Marvin J. [Bioorganic and Medicinal Chemistry, 2012, vol. 20, # 7, p. 2214 - 2220]

17
[ 51677-09-9 ]
[ 14441-90-8 ]

Yield	Reaction Conditions	Operation in experiment
79.5%	With sodium hydroxide; In methanol; at 20℃; for 2h;	To a solution of 1-2 (1.0 g, 4.9 mmol) in MeOH (10 mL) was added sodium hydroxide solution (20 mL, 4 M). The reaction mixture was stirred at room temperature for 2 hours and concentrated under reduced pressure to remove MeOH. The aqueous phase was acidified with aqueous HCl (1 M) till pH=3 and the mixture was extracted with EtOAc, dried with anhydrous Na2SO4, filtered and concentrated to give the crude product. The crude product was purified by silica gel chromatography eluted to give product 1-2 (0.7 g, 79.5%). MS m/z [ESI]: 190.0 [M+1].

Reference： [1]Indian Journal of Chemistry - Section B Organic and Medicinal Chemistry,2012,vol. 51,p. 1369 - 1375
[2]Patent: US2018/271846,2018,A1 .Location in patent: Paragraph 0159-0161
[3]Journal of Medicinal Chemistry,2018,vol. 61,p. 8337 - 8352
[4]Bioorganic and Medicinal Chemistry Letters,2013,vol. 23,p. 273 - 280

18
[ 14441-90-8 ]
[ 155252-35-0 ]
[ 1416981-44-6 ]

Yield	Reaction Conditions	Operation in experiment
89%	With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In dichloromethane at 0 - 20℃;	General procedure for the synthesis of compounds 16a-i, 17a-i and 18a-d General procedure: To a solution of compound 15 (1.0 mmol) in dichloromethane (20 ml) was added EDCI·HCl (1.2 mmol) and HOBt (0.1 mmol). Then added corresponding hetroaromatic acids 10a-i, 14a-i and 12a-d (1.0 mmol) and the reaction mixture was stirred at room temperature for 10 h. The reaction was monitored by TLC. After completion of reaction, water was added to reaction mixture and extracted with dichloromethane (2 × 30 ml). The solvent was evaporated under reduced pressure to afford the crude product which was further purified by column chromatography on silica gel using ethyl acetate and hexane as solvent system to obtain the pure products as solids. 4β-5-Phenylisoxazole-3-amido podophyllotoxin (16a): The compound 16a was prepared according to the general procedure. Yield 89%; mp: 155-156 °C; [α]25D -43.0 (c 0.5 in CHCl3), 1H NMR (300 MHz, CDCl3): δ 2.71-2.88 (m, 1H), 3.01 (dd, 1H, J = 3.7, 13.5 Hz), 3.75 (s, 6H), 3.78 (s, 3H), 3.84-3.94(m,1H), 4.38-4.57 (m, 2H), 5.40 (dd, 1H, J = 3.0, 7.7 Hz), 6.0 (d, 2H, J = 6.7 Hz), 6.23 (s, 2H), 6.48 (d, 1H, J = 15), 6.84 (s, 1H), 6.95 (s, 1H), 7.51 (m, 3H), 7.81 (dd, 2H, J = 2.2, 7.7 Hz); 13C NMR (300 MHz, CDCl3): δ 37.4, 41.7, 43.6, 46, 48.4, 56, 61.1, 68.9, 98.8, 101.7, 106.4, 108, 109.1, 109.9, 125.9, 126.4, 127.8, 129.3, 131.1, 132.3, 134.6, 137.1, 147.6, 152.7, 158.1, 158.7, 159.4, 174; MS (ESI): 585 [M+H]+; HRMS (ESI) Calcd for C32H29O9N2 [M+H]+ 585.1873. Found: 585.1865.

Reference： [1]Kamal, Ahmed; Tamboli, Jaki R.; Vishnuvardhan; Adil; Nayak, V. Lakshma; Ramakrishna [Bioorganic and Medicinal Chemistry Letters, 2013, vol. 23, # 1, p. 273 - 280]

19
[ 67-56-1 ]
[ 14441-90-8 ]
[ 51677-09-9 ]

Reference： [1]Synthesis,2013,vol. 45,p. 260 - 264

20
[ 14441-90-8 ]
[ 768-94-5 ]
[ 1009962-35-9 ]

Yield	Reaction Conditions	Operation in experiment
52%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In chloroform at 20℃; for 0.75h; Stage #2: 1-Adamantanamine In chloroform at 20℃; for 24h;	1 4.1.5. General procedure for the preparation of 5-arylisoxazole-3-carboxamides (44-70) General procedure: To a solution of carboxylic acid 33-43 in dry chloroform (20 mL) were added N,N-diisopropylethylamine (DIEA) (2 equiv) and 1-hydroxybenzotriazole (HOBt) (0.5 equiv), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) (1.5 equiv). The resulting mixture was stirred at room temperaturefor 45 min. The appropriate amine (1.2 equiv) was then added, and the solution was stirred at room temperature for additional 24 h. The solution was filtered and washed with 0.5 N aqueous NaOH (20 mL), with 1 N aqueous HCl (20 mL), and water (20 mL). The organic layer was dried over MgSO4 and evaporated under reduce pressure to give a brown oil. The crude material was purified by TLC using the appropriate eluent (cyclohexane/EtOAc 7:3, v/v) and recrystallized in heptane to afford the desired compounds.

Reference： [1]Tourteau, Aurélien; Andrzejak, Virginie; Body-Malapel, Mathilde; Lemaire, Lucas; Lemoine, Amélie; Mansouri, Roxane; Djouina, Madjid; Renault, Nicolas; El Bakali, Jamal; Desreumaux, Pierre; Muccioli, Giulio G.; Lambert, Didier M.; Chavatte, Philippe; Rigo, Benoît; Leleu-Chavain, Natascha; Millet, Régis [Bioorganic and Medicinal Chemistry, 2013, vol. 21, # 17, p. 5383 - 5394]

21
[ 14441-90-8 ]
[ 17768-41-1 ]
[ 1450606-88-8 ]

Yield	Reaction Conditions	Operation in experiment
34%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In chloroform at 20℃; for 0.75h; Stage #2: adamantylmethylamine In chloroform at 20℃; for 24h;	3 4.1.5. General procedure for the preparation of 5-arylisoxazole-3-carboxamides (44-70) General procedure: To a solution of carboxylic acid 33-43 in dry chloroform (20 mL) were added N,N-diisopropylethylamine (DIEA) (2 equiv) and 1-hydroxybenzotriazole (HOBt) (0.5 equiv), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) (1.5 equiv). The resulting mixture was stirred at room temperaturefor 45 min. The appropriate amine (1.2 equiv) was then added, and the solution was stirred at room temperature for additional 24 h. The solution was filtered and washed with 0.5 N aqueous NaOH (20 mL), with 1 N aqueous HCl (20 mL), and water (20 mL). The organic layer was dried over MgSO4 and evaporated under reduce pressure to give a brown oil. The crude material was purified by TLC using the appropriate eluent (cyclohexane/EtOAc 7:3, v/v) and recrystallized in heptane to afford the desired compounds.

Reference： [1]Tourteau, Aurélien; Andrzejak, Virginie; Body-Malapel, Mathilde; Lemaire, Lucas; Lemoine, Amélie; Mansouri, Roxane; Djouina, Madjid; Renault, Nicolas; El Bakali, Jamal; Desreumaux, Pierre; Muccioli, Giulio G.; Lambert, Didier M.; Chavatte, Philippe; Rigo, Benoît; Leleu-Chavain, Natascha; Millet, Régis [Bioorganic and Medicinal Chemistry, 2013, vol. 21, # 17, p. 5383 - 5394]

22
[ 14441-90-8 ]
[ 13074-39-0 ]
[ 1450606-87-7 ]

Yield	Reaction Conditions	Operation in experiment
51%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In chloroform at 20℃; for 0.75h; Stage #2: N-(2-adamantyl)amine In chloroform at 20℃; for 24h;	2 4.1.5. General procedure for the preparation of 5-arylisoxazole-3-carboxamides (44-70) General procedure: To a solution of carboxylic acid 33-43 in dry chloroform (20 mL) were added N,N-diisopropylethylamine (DIEA) (2 equiv) and 1-hydroxybenzotriazole (HOBt) (0.5 equiv), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) (1.5 equiv). The resulting mixture was stirred at room temperaturefor 45 min. The appropriate amine (1.2 equiv) was then added, and the solution was stirred at room temperature for additional 24 h. The solution was filtered and washed with 0.5 N aqueous NaOH (20 mL), with 1 N aqueous HCl (20 mL), and water (20 mL). The organic layer was dried over MgSO4 and evaporated under reduce pressure to give a brown oil. The crude material was purified by TLC using the appropriate eluent (cyclohexane/EtOAc 7:3, v/v) and recrystallized in heptane to afford the desired compounds.

Reference： [1]Tourteau, Aurélien; Andrzejak, Virginie; Body-Malapel, Mathilde; Lemaire, Lucas; Lemoine, Amélie; Mansouri, Roxane; Djouina, Madjid; Renault, Nicolas; El Bakali, Jamal; Desreumaux, Pierre; Muccioli, Giulio G.; Lambert, Didier M.; Chavatte, Philippe; Rigo, Benoît; Leleu-Chavain, Natascha; Millet, Régis [Bioorganic and Medicinal Chemistry, 2013, vol. 21, # 17, p. 5383 - 5394]

23
[ 144537-05-3 ]
[ 14441-90-8 ]

Reference： [1]Beilstein Journal of Organic Chemistry,2015,vol. 11,p. 1241 - 1245

24
[ 14441-90-8 ]
ethyl 5-(N-(2-aminoethyl)sulfamoyl)-1H-pyrazole-3-carboxylate hydrochloride [ No CAS ]
ethyl 5-(N-(2-(5-phenylisoxazole-3-carboxamido)ethyl)sulfamoyl)-1H-pyrazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
0.25 g	With 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane-2,4,6-trioxide; triethylamine In tetrahydrofuran at 20℃; for 6h;	4.4 Step-4: ethyl 5-(N-(2-(5-phenylisoxazole-3-carboxamido)ethyl)sulfamoyl)-lH- pyrazole-3-carboxylate a suspension of ethyl 5-(N-(2-aminoethyl)sulfamoyl)-lH-pyrazole-3- carboxylate (0.5 g), 5-phenylisoxazole-3-carboxylic acid (0.43 g, 0.00229 mol), Et3N (0.8 mL, 0.0057 mol) and T3P (1.8 g, 0.0057 mol) in 15 mL of anhydrous THF was stirred at room temperature. The reaction mixture was stirred at room temperature for 6 h. Volatiles were removed under vacuum and the crude was dissolved in ethyl acetate (20 mL). The organic layer was washed with saturated a2C03 solution (10 mL x 2), water, brine, dried over Na2S04 and concentrated under vacuum to obtain a residue which was purified by column (0219) chromatography on silica gel (100-200 mesh) using 5% methanol in DCM to afford product (0.250 g, 35% from step 2) as off-white solid. XH NMR (400 MHz,CDCl3+2drops DMSO-d6): δ 7.88 (br, 1H), 7.77-7.75 (m, 2H), 7.46-7.45 (m, 3H), 7.13 (s, 1H), 6.91 (s, 1H), 6.65 (br, 1H), 4.30 (q, J = 1.1 Hz, 2H), 3.59-3.55 (m, 2H), 3.35-3.31 (m, 2H), 1.32 (t, J = 1.1 Hz, 3H); LC- MS: [M+H]+ = 434.2 m/z.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/138934, 2015, A1 Location in patent: Paragraph 00137

25
[ 14441-90-8 ]
[ 75178-96-0 ]
tert-butyl N-[3-[(5-phenyl-1,2-oxazol-3-yl)formamido]propyl]carbamate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
85%	With 1-[(1-(cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino)]-uronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide	13.i Preparation of TERT-BUTYL (3-(5-PHENYLISOXAZOLE-3-CARBOXAMIDO)PROPYL)CARBAMATE To a solution containing 1.0 g (5.29 mmol) of 3-phenylisoxazole-3-carboxylic acid and 0.89 g (5.56 mmol) of tert-butyl (3-aminopropyl)carbamate in 10 mL of DMF was added 2.5 g (5.82 mmol) of COMU, followed by 2.0 mL (11.1 mmol) of DIPEA. The reaction mixture was allowed to stir at rt overnight. The solvents were removed under reduced pressure and the residue was subjected to silica gel chromatography to give 1.55 g (85%) of tert-butyl (3-(5-phenylisoxazole-3-carboxamido)propyl)carbamate as a yellow solid. LC/MS: 1.22 min, m/z=368.2 [M+K]+
85%	With 1-[(1-(cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino)]-uronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃;	1.i i) Preparation of tert-butyl (3-(5-phenylisoxazole-3-carboxamido)propyl)carbamate (0406) To a solution containing 1.0 g (5.29 mmol) of 3-phenylisoxazole-3-carboxylic acid and 0.89 g (5.56 mmol) of tert-butyl(3-aminopropyl)carbamate in 10 mL of DMF was added 2.5 g (5.82 mmol) of COMU, followed by 2.0 mL (11.1 mmol) of DIPEA. The reaction mixture was allowed to stir at rt overnight. The solvents were removed under reduced pressure and the residue was subjected to silica gel chromatography to give 1.55 g (85%) of tert-butyl (3-(5-phenylisoxazole-3-carboxamido)propyl)carbamate as a yellow solid: LC/MS: 1.22 min, m/z=368.2 [M+K]+.
41%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	6.1 Step 1: tert-butyl N-[3-[(5-phenyl-l,2-oxazol-3- yl)formamido]propyl]carbamate tert-butyl N-(3-aminopropyl)carbamate (884 mg, 5.07 mmol, 1.20 eq.), HATU (1930 mg, 5.08 mmol, 1.20 eq.) were added to a solution of 5-phenyl- l,2-oxazole-3-carboxylic acid (800 mg, 4.23 mmol, 1.00 eq.) in dichloromethane (50 mL). DIEA (1638 mg, 12.67 mmol, 3.00 eq.) was added dropwise to the reaction mixture and then it was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water, extracted with dichloromethane (3x50 mL) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with dichloromethane/methanol (100: 1) to give 600 mg (41%) of tert-butyl N-[3-[(5-phenyl-l,2-oxazol-3-yl)formamido]propyl]carbamate as a white solid. LC-MS: (0242) [M+H]+=346.

Reference： [1]Current Patent Assignee: VANDERBILT UNIVERSITY - US2016/52895, 2016, A1 Location in patent: Paragraph 0465; 0466
[2]Current Patent Assignee: VANDERBILT UNIVERSITY - US2016/52896, 2016, A1 Location in patent: Paragraph 0405-0406
[3]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/138934, 2015, A1 Location in patent: Paragraph 00155

26
[ 14441-90-8 ]
ethyl 5-(N-(3-aminopropyl)sulfamoyl)-1H-pyrazole-3-carboxylate hydrochloride [ No CAS ]
ethyl 5-(N-(3-(5-phenylisoxazole-3-carboxamido)propyl)sulfamoyl)-1H-pyrazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
0.45 g	With 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane-2,4,6-trioxide; triethylamine In tetrahydrofuran at 20℃; for 6h;	5.3 Step-3: ethyl 5-(N-(3-(5-phenylisoxazole-3-carboxamido)propyl)sulfamoyl)-lH- pyrazole-3-carboxylate: a suspension of ethyl 5-(N-(3-aminopropyl)sulfamoyl)-lH-pyrazole- 3-carboxylate (0.6 g), 5-phenylisoxazole-3-carboxylic acid (0.493 g, 0.0026 mol), Et3N (1.1 niL, 0.0065 mol) and T3P (2 g, 0.0065 mol) in anhydrous THF (20 mL) was stirred at room temperature for 6 h. Volatiles were removed under reduced pressure and the residue was dissolved in ethyl acetate (20 mL). The organic layer was washed with saturated a2C03 solution (10 mL x 2), water, brine, dried over Na2S04 and concentrated under reduced pressure to afford obtain crude compound. The crude was purified by column chromatography on silica gel (100-200 mesh) using 5% methanol in DCM to give product (0.450 g, 63 %) as off-white solid. 'H NMR (400 MHz, CDC13): δ 11.91 (br, 1H), 7.79-7.77 (m, 2H), 7.50-7.46 (m, 3H), 7.37 (m, 1H), 7.24 (s, 1H), 6.98 (s, 1H), 4.41-4.36 (q, J= 7.1 Hz, 2H), 3.62-3.57 (m, 2H), 3.26- 21 (m, 2H), 1.89-1.84 (m, 2H), 1.37 (t, J= 7.1 Hz, 3H); LC-MS: [M+H]+ = 448.0 m/z.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/138934, 2015, A1 Location in patent: Paragraph 00146

27
[ 14441-90-8 ]
[ 107-35-7 ]
2-(5-phenylisoxazole-3-carboxamido)ethane-1-sulfonic acid [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
10%	With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In water; N,N-dimethyl-formamide at 20℃; for 72h;	8.1 Step 1: 2-(5-phenylisoxazole-3-carboxamido)ethane-l-sulfonic acid: HOBt (2.56 g, 0.019 mol) was added to a solution of 2-aminoethane-l -sulfonic acid (2.97 g, 0.024 mol) and 5-phenylisoxazole-3-carboxylic acid (3.0 g, 0.016 mol) in DMF (150 mL) and water (25 mL), followed by EDC HCl (4.54 g, 0.024 mol). Then DIPEA (6.12 g, 0.047 mol) was added drop wise to the reaction mixture and stirred at room temperature for 72 h. Volatiles were removed under reduced pressure to afford crude compound, which was further purified by preparative HPLC to obtain the product (500 mg, 10%) as off white solid. NMR (400 MHz, D20): δ 7.81-7.79 (m, 2H), 7.52-7.51 (m, 3H), 6.94 (s, 1H), 3.73 (t, J=6.9 Hz, 2H), 3.17 (t, J=6.8 Hz, 2H), LC-MS: [M-H]"=295.1.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/138934, 2015, A1 Location in patent: Paragraph 00165

28
[ 14441-90-8 ]
[ 150349-36-3 ]
tert-butyl methyl(3-(5-phenylisoxazole-3-carboxamido)propyl)carbamate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
36.74%	With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; In tetrahydrofuran; at 20℃; for 16h;	EDC HC1 (0.30 g, 0.0015 mol) was added to a solution of 5- phenylisoxazole-3-carboxylic acid (0.2 g, 0.0010 mol) and tert-butyl (3- aminopropyl)(methyl)carbamate (0.19 g, 0.0010 mol) in THF (4 mL) followed by addition HOBt (0.21 g, 0.0015 mol) at rt and reaction mixture was stirred at rt for 16 h. Progress the reaction was monitored by TLC. After completion, reaction was quenched with water (20 mL), extracted with ethyl acetate (2 x 25 mL), combined organic layer was dried over sodium sulfate and evaporated volatiles under vacuum. The crude was purified by column chromatography using 100-200 mesh silica eluted with 50 % ethyl acetate in Hexane to give product (0.14 g, 36.74%) as a white solid. NMR (400 MHz, CDC13): delta 7.79-7.77 (m, 2H), 7.47-7.46 (m, (0191) 3H), 6.94 (s, 1H), 3.44 (br, 2H), 3.35 (br, 2H), 2.85 (s, 3H), 1.79 (br, 2H), 1.46 (s, 9H), LC-MS: [M+H]+ 360.1.

Reference： [1]Patent: WO2015/138934,2015,A1 .Location in patent: Paragraph 00114

29
[ 14441-90-8 ]
[ 57260-73-8 ]
tert-butyl(2-(5-phenylisoxazole-3-carboxamido)ethyl)carbamate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
71%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With 1,1'-carbonyldiimidazole In dichloromethane at 20℃; for 0.25h; Stage #2: N-BOC-1,2-diaminoethane With N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃;	1.i i) Preparation of TERT-BUTYL (2-(5-PHENYL- ISOXAZOLE-3-CARBOXAMIDO)ETHYE)CARBAMATE To a solution containing 1.0 g (5.29 mmol) of 5-phenylisoxazole-3-carboxylic acid and 15 mE of DCM was added 0.943 g (5.82 mmol) of CDI. The reaction mixture was allowed to stir at it for 15 mm and a solution containing 0.889 g (5.56 mmol) of tert-butyl (2-aminoethyl)carbamate inS mE of DCM was added, followed by 2 mE (11 mmol) of DIPEA. The reaction mixture was allowed to stir at it overnight, quenched by the addition of water, and extracted with DCM. The combined organic layers were dried by passage through a phase separator cartridge and the solvent was removed under reduced pressure. The residue was subjected to silica gel chromatography to give 1.21 g (7 1%) of tert-butyl (2-(5- phenylisoxazole-3 -carboxamido)ethyl)carbamate as a white solid. LC/MS ret time: 1.201 min m/z=276.2 [M-C(CH3)3)]+

Reference： [1]Current Patent Assignee: VANDERBILT UNIVERSITY - US2016/52895, 2016, A1 Location in patent: Paragraph 0438; 0439

30
[ 14441-90-8 ]
[ 4138-35-6 ]
methyl 3-(5-phenylisoxazole-3-carboxamido)propanoate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
6.5 g	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 0℃; for 0.5h; Stage #2: methyl 3-aminopropanoate In N,N-dimethyl-formamide for 0.166667h; Stage #3: With N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 25℃; for 1h;	To a stirred solution of 5- phenylisoxazole-3-carboxylic acid (5.0g, 26.43mmol) in DMF (50 mL) was added HATU (15.06 g, 39.63mmol) at 0°C. The reaction mixture was stirred at 0°C for 30 minutes and then methyl 3-aminopropanoate (3.65g, 26.43mmol) was added. After stirring 10 minutes, DIPEA (24 mL, 132.15mmol) was added drop wise and the reaction stirred at 25°C for lh. The reaction mixture was quenched in ice cold water and the resulting solid was filtered. The crude product was purified by column chromatography (10-15% ethyl acetate in hexane) to give methyl 3-(5-phenylisoxazole-3-carboxamido) propanoate (6.5g, 275 [M+H]). 1H NMR: (400MHz, DMSO): δ: 2.605-2.641 (t, 2H), 3.062 (s, 2H), 3.618 (s, 3H), 7.539-7.609 (m, 4H), 7.930-7.954 (dd, J=1.6, 7.2, 2H), 8.875-8.903 (t, 1H).

Reference： [1]Current Patent Assignee: FLATLEY DISCOVERY LAB LLC - WO2016/54560, 2016, A1 Location in patent: Page/Page column 39

31
[ 14441-90-8 ]
5-aminovaleric acid methyl ester hydrochloride [ No CAS ]
[ 1045792-81-1 ]

Yield	Reaction Conditions	Operation in experiment
9.5 g	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 0℃; for 0.5h; Stage #2: 5-aminovaleric acid methyl ester hydrochloride In N,N-dimethyl-formamide for 0.25h; Stage #3: With N-ethyl-N,N-diisopropylamine at 25℃; for 2h;	5-(5-phenylisoxazole-3-carboxamido)pentanoate To a stirred solution of 5- aminopentanoic acid (7.35 g, 59.8mmol) in methanol (50mL) was added thionyl chloride (4.9mL,65.8mmol) drop wise at 0°C. The reaction was stirred at 25°C for 16h. The reaction mixture was concentrated under reduce pressure and the resulting residue was suspended in diethyl ether (lOOmL). The solid precipitate was collected by filtration and dried under vacuum to obtain methyl 5-aminopentanoate hydrochloride. To a stirred solution of 5- phenylisoxazole-3-carboxylic acid (7.4 g, 39. lmmol) in DMF (80mL) was added HATU (22.3g, 58.6mmol) at 0°C and the reaction mixture was then stirred at 0°C for 30 minutes. Methyl 5-aminopentanoate hydrochloride (7.8g, 46.9mmol) was added portion wise and the reaction mixture was stirred for 15 minutes. DIPEA (34.0mL, 195mmol) was added drop wise and the reaction mixture was stirred at 25°C for 2h. Reaction mixture was diluted with ice water (500mL) and the solid precipitate was collected by filtration and dried under vacuum to obtained methyl 5-(5-phenylisoxazole-3-carboxamido)pentanoate (9.5 g, 279 [M+H]); 1H MR: (400 MHz, DMSO) δ: 1.545-1.580(m, 4H), 2.336-2.371 (m, 2H), 3.243- 3.289(d, J=12.4, 2H), 3.589(S, 3H), 7.364 (S, 1H), 7.538-7.581 (m, 3H), 7.927-7.951 (dd,J=1.6, 7.2, 2H), 8.836-8.865 (t, lH).

Reference： [1]Current Patent Assignee: FLATLEY DISCOVERY LAB LLC - WO2016/54560, 2016, A1 Location in patent: Page/Page column 36; 37

32
[ 14441-90-8 ]
5-amino-1-(4-methylpiperazin-1-yl)pentan-1-one [ No CAS ]
N-(5-(4-methylpiperazin-1-yl)-5-oxopentyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 0℃; for 0.5h; Stage #2: 5-amino-1-(4-methylpiperazin-1-yl)pentan-1-one In N,N-dimethyl-formamide at 0℃; for 0.25h; Stage #3: With N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 0 - 25℃; for 1h;	43 N-(5-(4-methylpiperazin-l-yl)-5-oxopentyl)-5-phenylisoxazole-3- carboxamide To a stirred solution of 5-phenylisoxazole-3-carboxylic acid (0.5 (0229) g,2.64mmol) in DMF (lOmL) was added HATU (1.5g, 3.96mmol) at 0°C. The reaction was stirred at 0°C for 30min. 5-amino-l-(4-methylpiperazin-l-yl)pentan-l-one (0.526g, 2.64mmol) was added at 0°C and the reaction was stirred for 15 min. Next, DIPEA (3.2mL, 18.4mmol) was added at 0°C and the reaction was stirred at 25°C for lh. Water was added (lOOmL) and the resulting solid was filtered and dried under vacuum to give N-(5-(4- methylpiperazin-l-yl)-5-oxopentyl)-5-phenylisoxazole-3-carboxamide (0.120g, 371 [M+H]). 1H NMR: (400MHz, DMSO) : δ: 1.535-1.543 (d, J=3.2, 4H), 2.202 (s, 3H), 2.281 (s, 2H), 2.335 (s, 4H), 3.262-3.278 (d, J=6.4, 2H), 3.439 (s, 4H), 7.364 (s, 1H), 7.553-7.568 (d, J=6, 3H), 7.931-7.950 (t, 2H), 8.844-8.857 (d, J=5.2, 1H).

Reference： [1]Current Patent Assignee: FLATLEY DISCOVERY LAB LLC - WO2016/54560, 2016, A1 Location in patent: Page/Page column 38; 39

33
[ 14441-90-8 ]
C₂₈H₄₅FN₄O₄ [ No CAS ]
C₃₃H₄₂FN₅O₄ [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	With N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 0 - 20℃;	64-a Step 1: To a solution of intermediate 12-g (847 g, 1.62 mmol) in DMF, cooled to 0° C., were sequentially added 5-phenylisoxazole-3-carboxylic acid (431 mg, 2.27 mmol), HATU (1.05 g, 2.77 mmol) and DIPEA (1.13 mL, 6.51 mmol) and the reaction mixture was stirred at room temperature overnight. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride, saturated aqueous NaHCO3 and brine, dried over anhydrous MgSO4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 64-a as a white foam.

Reference： [1]Current Patent Assignee: PHARMASCIENCE INC - US9284350, 2016, B2 Location in patent: Page/Page column 137

34
[ 14441-90-8 ]
tert-butyl N-(3-azidocyclohexyl)carbamate [ No CAS ]
[ 107-19-7 ]
N-[3-[5-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclohexyl]-5-phenylisoxazole-3-carboxamide [ No CAS ]
N-[3-[4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclohexyl]-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 5% 2: 6%	Stage #1: tert-butyl N-(3-azidocyclohexyl)carbamate; propargyl alcohol In N,N-dimethyl-formamide at 100℃; Sealed tube; Stage #2: With hydrogenchloride In 1,4-dioxane; water at 20℃; for 4h; Stage #3: 5-phenyl-3-isoxazolecarboxylic acid With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 5h;	64.4 [0647] Step 4: N-j3- 15-(hydroxymethyl)-1H-1,2,3-triazol-1-ylj cyclohexylj -5- phenylisoxazole-3-carboxamide and N-13-14-(hydroxymethyl)-1H-1,2,3-triazol-1- yljcyclohexylj-5-phenylisoxazole-3-carboxamide: a solution of tert-butyl N-(3-azidocyclohexyl)carbamate (642 mg, 2.67 mmol, 1.00 eq.) and prop-2-yn-1-ol (300 mg, 5.35 mmol, 2.00 eq.) in DMF (10 mL) wasplaced in a sealed tube and the solution was stirred for overnight at 100°C in an oil bath. The resulting mixture was concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (3:1). This resulted in 582 mg (crude) mixture of tert-butyl N-[3 - [4/5-(hydroxymethyl)- 1 H- 1,2,3 -triazol- 1 -yl]cyclohexyl]carbamate.[hydrogen chloride (3 mL) was added to a solution of mixture of tert-butyl N-[3-[4/5-(hydroxymethyl)- 1H-1,2,3-triazol-1-yl]cyclohexyl]carbamate (582 mg, 1.96 mmol, 1.00 eq., crude) in 1,4- dioxane (10 mL). The resulting solution was stirred for 4 hours at room temperature and then it was concentrated under vacuum. This resulted in 637 mg of mixture of [1-(3-aminocyclohexyl)-1H-1,2,3-triazol-4/5-yl]methanol as a brown oilDIEA (1.26 g, 3.00 eq.), HATU (2.47 g,2.00 eq.) and 5-phenylisoxazole-3-carboxylic acid (1.23 g, 6.5 mmol, 2.00 eq.) were added to a mixture of [1 -(3 -aminocyclohexyl)- 1 H- 1,2,3 -triazol-5 -yl]methanol and [1 -(3- aminocyclohexyl)-1H-1,2,3-triazol-4-yl]methanol (637 mg, 3.25 mmol, 1.00 eq.) in dichloromethane (100 mL). The resulting solution was stirred for 5 hours at room temperature and the reaction was then quenched by the addition of 40 mL of water. The resulting solutionwas extracted with dichloromethane (3x100 mL) and the combined organic layers were washed with brine (2x20 mL), dried over anhydrous sodium sulfate and concentrated under vacuum.The residue was applied onto a silica gel column with ethyl acetate (100%). This resulted in25.9 mg (6%) of N-[3-[5-(hydroxymethyl)- 1H- 1 ,2,3-triazol- 1 -yl]cyclohexyl]-5-phenylisoxazole-3-carboxamide as an off-white solid and 36.4 mg (5%) of N-[3-[4-(hydroxymethyl)- 1 H- 1,2,3 -triazol- 1 -yl]cyclohexyl]-5-phenylisoxazole-3 -carboxamide as anwhite solid.[0648] N- 13-14-(hydroxymethyl)-1H-1,2,3-triazol-1-ylj cyclohexyll -5-phenylisoxazole-3- carboxamide:[0649] Analytical data:[0650] LC-MS (ES, m/z): [M+H] = 368.1[0651] ‘HNMR (DM50-cl6, 400 MHz): ö 8.82-8.80 (d, J = 7.6 Hz, 1H), 8.05 (s, 1H), 7.95-7.93 (m, 2H), 7.58-7.54 (m, 3H), 7.37 (s, 1H), 5.18-5.15 (m, 1H), 4.93 (br, 1H),4.53-4.50 (m, 2H), 4.3 1-4.25 (m, 1H), 2.38-2.30 (m, 1H), 2.18-2.11 (m, 1H),1.97-1.95 (m, 2H), 1.73-1.7 1 (m, 4H).[0652] N- 13-15-(hydroxymethyl)-1H-1,2,3-triazol-1-ylj cyclohexyll -5-phenylisoxazole-3-carboxamide:[0653] Analytical data:[0654] LC-MS (ES, m/z): [M+H] = 368.1[0655] ‘H NMR (DMSO-d6, 400 MHz): ö 8.77-8.75 (d, J= 7.6 Hz, 1H), 7.94-7.93 (m, 2H), 7.60-7.55 (m, 4H), 7.37 (s, 1H), 5.50-5.43 (m, 1H), 4.92-4.91 (m, 1H), 4.60-4.52(m, 3H), 2.37-2.12 (m, 2H), 2.03-1.82 (m, 2H), 1.75 (br, 3H), 1.60-1.45 (m, 1H),1.82-1.75 (m, 4H).
1: 25.9 mg 2: 36.4 mg	Stage #1: tert-butyl N-(3-azidocyclohexyl)carbamate; propargyl alcohol In N,N-dimethyl-formamide at 100℃; Sealed tube; Stage #2: With hydrogenchloride In 1,4-dioxane at 20℃; for 4h; Stage #3: 5-phenyl-3-isoxazolecarboxylic acid With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 5h;	64.4 N-[3-[5-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclohexyl]-5-phenylisoxazole-3-carboxamide and N-[3-[4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclohexyl]-5-phenylisoxazole-3-carboxamide a solution of tert-butyl N-(3-azidocyclohexyl)carbamate (642 mg, 2.67 mmol, 1.00 eq.) and prop-2-yn-l-ol (300 mg, 5.35 mmol, 2.00 eq.) in DMF (10 mL) was placed in a sealed tube and the solution was stirred for overnight at 100°C in an oil bath. The resulting mixture was concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (3: 1). This resulted in 582 mg (crude) mixture of tert-butyl N-[3-[4/5-(hydroxymethyl)-lH-l,2,3-triazol-l-yl]cyclohexyl]carbamate. LC-MS (ES, m/z): [M+H]+ = 297.2; hydrogen chloride (3 mL) was added to a solution of mixture of tert-butyl N-[3-[4/5-(hydroxymethyl)- lH-l,2,3-triazol-l-yl]cyclohexyl]carbamate (582 mg, 1.96 mmol, 1.00 eq., crude) in 1,4- dioxane (10 mL). The resulting solution was stirred for 4 hours at room temperature and then it was concentrated under vacuum. This resulted in 637 mg of mixture of [l-(3- aminocyclohexyl)-lH-l,2,3-triazol-4/5-yl]methanol as a brown oil. LC-MS (ES, m/z): [M+H]+ = 197.1; DIEA (1.26 g, 3.00 eq ), HATU (2.47 g, 2.00 eq.) and 5-phenylisoxazole-3-carboxylic acid (1.23 g, 6.5 mmol, 2.00 eq.) were added to a mixture of [l-(3-aminocyclohexyl)-lH-l,2,3-triazol-5-yl]methanol and [l-(3- aminocyclohexyl)-lH-l,2,3-triazol-4-yl]methanol (637 mg, 3.25 mmol, 1.00 eq.) in dichloromethane (100 mL). The resulting solution was stirred for 5 hours at room temperature and the reaction was then quenched by the addition of 40 mL of water. The resulting solution was extracted with dichloromethane (3x100 mL) and the combined organic layers were washed with brine (2x20 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate (100%). This resulted in 25.9 mg (6%) of N-[3-[5-(hydroxymethyl)-lH-l,2,3-triazol-l-yl]cyclohexyl]-5- phenylisoxazole-3-carboxamide as an off-white solid and 36.4 mg (5%) of N-[3-[4- (hydroxymethyl)-lH-l,2,3-triazol-l-yl]cyclohexyl]-5-phenylisoxazole-3-carboxarnide as an white solid. A^-[3-[4-(hydroxymethyl)-lH-l,2,3-triazol-l-yl]cyclohexyl]-5-phenylisoxazole-3- carboxamide: [0597] Analytical data: [0598] LC-MS (ES, m/z): [M+H]+ = 368.1 [0599] XH NMR (DMSO-i, 400 MHz): δ 8.82-8.80 (d, J = 7.6 Hz, 1H), 8.05 (s, 1H), 7.95- 7.93 (m, 2H), 7.58-7.54 (m, 3H), 7.37 (s, 1H), 5.18-5.15 (m, 1H), 4.93 (br, 1H), 4.53-4.50 (m, 2H), 4.31-4.25 (m, 1H), 2.38-2.30 (m, 1H), 2.18-2.11 (m, 1H), 1.97-1.95 (m, 2H), 1.73-1.71 (m, 4H). A^-[3-[5-(hydroxymethyl)-lH-l,2,3-triazol-l-yl]cyclohexyl]-5-phenylisoxazole-3- carboxamide: [0600] Analytical data: [0601] LC-MS (ES, m/z): [M+H]+ = 368.1 [0602] lH NMR (DMSO-i, 400 MHz): δ 8.77-8.75 (d, J= 7.6 Hz, 1H), 7.94-7.93 (m, 2H), 7.60-7.55 (m, 4H), 7.37 (s, 1H), 5.50-5.43 (m, 1H), 4.92-4.91 (m, 1H), 4.60-4.52 (m, 3H), 2.37- 2.12 (m, 2H), 2.03-1.82 (m, 2H), 1.75 (br, 3H), 1.60-1.45 (m, 1H), 1.82-1.75 (m, 4H).

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0645; 0646; 0647-0655
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0594-0602

35
[ 14441-90-8 ]
tert-butyl N-(3-azidocyclopentyl)carbamate [ No CAS ]
[ 107-19-7 ]
N-[3-[5-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclopentyl]-5-phenylisoxazole-3-carboxamide [ No CAS ]
N-[3-[4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclopentyl]-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 54.2 mg 2: 124.4 mg	Stage #1: tert-butyl N-(3-azidocyclopentyl)carbamate; propargyl alcohol In N,N-dimethyl-formamide at 100℃; for 16h; Stage #2: With hydrogenchloride In 1,4-dioxane; water at 25℃; for 3h; Stage #3: 5-phenyl-3-isoxazolecarboxylic acid With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 25℃; for 3h;	69.4 [0702] Step 4: N-j3- 15-(hydroxymethyl)-1H-1,2,3-triazol-1-ylj cyclopentylj -5- phenylisoxazole-3-carboxamide and N-13-14-(hydroxymethyl)-1H-1,2,3-triazol-1- ylj cyclopentylj-5-phenylisoxazole-3-carboxamide: a solution of tert-butyl N-(3-azidocyclopentyl)carbamate (785 mg, 3.47 mmol, 1.00 eq.) and prop-2-yn-1-ol (387 mg, 6.90 mmol, 2.00 eq.) in DMF (5 mL) wasstirred for 16 hours at 100°C. The resulting mixture was concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (2:1) to give 620 mg (63%) of a mixture of tert-butyl N- [3- [5-(hydroxymethyl)- 1 H- 1,2,3 -triazol- 1- yl]cyclopentyl]carbamate and tert-butyl N-[3 -[4-(hydroxymethyl)- 1 H- 1,2,3 -triazol- 1- yl]cyclopentyl]carbamate as a yellow oil.concentrated hydrogen chloride (3 mL)was added to a solution of the mixture of tert-butyl N-[3-[5-(hydroxymethyl)-1H-1,2,3-triazol-1 -yl]cyclopentyl]carbamate and tert-butyl N- [3- [4-(hydroxymethyl)- 1 H- 1,2,3 -triazol- 1- yl]cyclopentyl]carbamate (620 mg, 2.20 mmol, 1.00 eq.) in 1,4-dioxane (10 mL) and the mixture was stirred for 3 hours at 25°C. The mixture was concentrated under vacuum. This resulted in 600 mg (crude) of a mixture of [1-(3-aminocyclopentyl)-1H-1,2,3-triazol-5-yl]methanol and [1 -(3 -aminocyclopentyl)- 1 H- 1,2,3 -triazol-4-yl]methanol as yellow oil.[1 -(3 -aminocyclopentyl)- 1 H- 1,2,3 -triazol-5 -yl]methanol and [1 -(3 -aminocyclopentyl)- 1 H- 1,2,3 -triazol-4-yl]methanol (327 mg,1.8 mmol, 1.20 eq.) were added to a solution of 5-phenylisoxazole-3-carboxylic acid (285 mg,1.50 mmol, 1.00 eq.), HATU (855 mg, 2.25 mmol, 1.50 eq.) and DIEA (580 mg, 4.49 mmol,3.00 eq.) in dichloromethane (10 mL) and the mixture was stirred for 3 hours at 25°C. Theresulting solution was diluted with 100 mL of H20, extracted with ethyl acetate (2x 100 mL)and the organic layers combined. The resulting mixture was washed with brine (2x100 mL),dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product was purified by Prep-TLC (petroleum ether/ethyl acetate=1 :2). This resulted in 54.2 mg (10%) of N-[3 - [5-(hydroxymethyl)- 1 H- 1,2,3 -triazol- 1 -yl]cyclopentyl] -5 -phenylisoxazole-3 -carboxamide as a light yellow solid and 124.4 mg (23%) of N-[3-[4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclopentyl]-5-phenylisoxazole-3 -carboxamide as a white solid.[0703] Analytical data:[0704] N- 13-15-(hydroxymethyl)-1H-1,2,3-triazol-1-ylj cyclopentylj-5-phenylisoxazole- 3-carboxamide[0705] LC-MS (ES, m/z): [M+H] = 354.3 ‘H NMR (400 MHz, DMSO-d6) ö 9.03 -9.02 (d, J= 7.6 Hz, 1H), 7.95-7.92 (m, 2H),7.59-7.52 (m, 4H), 7.38 (s, 1H), 5.48-5.45 (t, J= 5.6 Hz, 1H), 5.03-4.96 (m,1H), 4.62-4.60 (d, J= 5.6 Hz, 2H), 4.49-4.43 (q, J= 7.6 Hz, 1H), 2.60-2.57 (m,1H), 2.26-2.2 1 (m, 3H), 2. 14-2.04 (m, 1H), 2.00-1.93 (m, 1H).N- 13-14-(hydroxymethyl)-1H-1,2,3-triazol-1-ylj cyclopentylj-5-phenylisoxazole-3-carboxamideLC-MS (ES, m/z): [M+H] = 354.3‘HNMR (400 MHz, DMSO-d6) ö 9.06 (d, J= 7.8 Hz, 1H), 8.10 (s, 1H), 7.97- 7.92 (m, 2H), 7.63 - 7.52 (m, 3H), 7.39 (s, 1H), 5.19 (t, J= 5.6 Hz, 1H), 5.03 (q,J= 7.6 Hz, 1H), 4.53 (d, J= 5.7 Hz, 2H), 4.47 (q, J= 7.6 Hz, 1H), 2.64 (dd, J=14.3, 6.7 Hz, 1H), 2.29 - 1.86 (m, 5H).
1: 54.2 mg 2: 124.4 mg	Stage #1: tert-butyl N-(3-azidocyclopentyl)carbamate; propargyl alcohol In N,N-dimethyl-formamide at 100℃; for 16h; Stage #2: With hydrogenchloride In 1,4-dioxane at 25℃; for 3h; Stage #3: 5-phenyl-3-isoxazolecarboxylic acid With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 25℃; for 3h;	69.4 N-[3-[5-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclopentyl]-5-phenylisoxazole-3-carboxamide and N-[3-[4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]cyclopentyl]-5-phenylisoxazole-3-carboxamide a solution of tert-butyl N-(3-azidocyclopentyl)carbamate (785 mg, 3.47 mmol, 1.00 eq.) and prop-2-yn-l-ol (387 mg, 6.90 mmol, 2.00 eq.) in DMF (5 mL) was stirred for 16 hours at 100°C. The resulting mixture was concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (2: 1) to give 620 mg (63%) of a mixture of tert-butyl N-[3-[5-(hydroxymethyl)-lH-l,2,3-triazol-l- yl]cyclopentyl]carbamate and tert-butyl N-[3-[4-(hydroxymethyl)-lH-l,2,3-triazol-l- yl]cyclopentyl] carbamate as a yellow oil. LC-MS (ES, m/z): [M+H]+ = 283.2; concentrated hydrogen chloride (3 mL) was added to a solution of the mixture of tert-butyl N-[3-[5-(hydroxymethyl)-lH-l,2,3-triazol- l-yl]cyclopentyl]carbamate and tert-butyl N-[3-[4-(hydroxymethyl)-lH-l,2,3-triazol-l- yl]cyclopentyl]carbamate (620 mg, 2.20 mmol, 1.00 eq.) in 1,4-dioxane (10 mL) and the mixture was stirred for 3 hours at 25°C. The mixture was concentrated under vacuum. This resulted in 600 mg (crude) of a mixture of [l-(3-aminocyclopentyl)-lH-l,2,3-triazol-5- yljmethanol and [l-(3-aminocyclopentyl)-lH-l,2,3-triazol-4-yl]methanol as yellow oil. LC- MS (ES, m/z): [M+H]+ = 183.1; [l-(3-aminocyclopentyl)-lH-l,2,3-triazol- 5-yl]methanol and [l-(3-aminocyclopentyl)-lH-l,2,3-triazol-4-yl]methanol (327 mg, 1.8 mmol, 1.20 eq.) were added to a solution of 5-phenylisoxazole-3-carboxylic acid (285 mg, 1.50 mmol, 1.00 eq.), HATU (855 mg, 2.25 mmol, 1.50 eq.) and DIEA (580 mg, 4.49 mmol, 3.00 eq.) in dichloromethane (10 mL) and the mixture was stirred for 3 hours at 25°C. The resulting solution was diluted with 100 mL of H20, extracted with ethyl acetate (2x100 mL) and the organic layers combined. The resulting mixture was washed with brine (2x100 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product was purified by Prep-TLC (petroleum ether/ethyl acetate=l :2). This resulted in 54.2 mg (10%) of N-[3-[5- (hydroxymethyl)-lH-l,2,3 riazol-l-yl]cyclopentyl]-5-phenylisoxazole-3-carboxarnide as a light yellow solid and 124.4 mg (23%) of N-[3-[4-(hydroxymethyl)-lH-l,2,3-triazol-l- yl]cyclopentyl]-5-phenylisoxazole-3-carboxamide as a white solid. [0648] Analytical data: N- [3- [5-(hyd roxymethyl)- IH- 1,2,3-triazol- 1-yl] cyclopentyl] -5-phenylisoxazole-3- carboxamide [0649] LC-MS (ES, m/z): [M+H]+ = 354.3 [0650] XH NMR (400 MHz, DMSO-c) δ 9.03-9.02 (d, J = 7.6 Hz, IH), 7.95-7.92 (m, 2H), 7.59-7.52 (m, 4H), 7.38 (s, IH), 5.48-5.45 (t, J = 5.6 Hz, IH), 5.03-4.96 (m, IH), 4.62-4.60 (d, J = 5.6 Hz, 2H), 4.49-4.43 (q, J = 7.6 Hz, IH), 2.60-2.57 (m, IH), 2.26-2.21 (m, 3H), 2.14-2.04 (m, IH), 2.00-1.93 (m, IH). N- [3- [4-(hydroxymethyl)- IH- 1,2,3-triazol- 1-yl] cyclopentyl] -5-phenylisoxazole-3- carboxamide [0651] LC-MS (ES, m/z): [M+H]+ = 354.3 [0652] XH NMR (400 MHz, DMSO-c) δ 9.06 (d, J = 7.8 Hz, IH), 8.10 (s, IH), 7.97 - 7.92 (m, 2H), 7.63 - 7.52 (m, 3H), 7.39 (s, IH), 5.19 (t, J = 5.6 Hz, IH), 5.03 (q, J = 7.6 Hz, IH), 4.53 (d, J = 5.7 Hz, 2H), 4.47 (q, J = 7.6 Hz, IH), 2.64 (dd, J = 14.3, 6.7 Hz, IH), 2.29 - 1.86 (m, 5H).

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0700; 0701; 0702-0709
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0645-0647

36
[ 14441-90-8 ]
1-[1-[trans-3-aminocyclobutyl]-1H-pyrazol-3-yl]ethan-1-ol hydrochloride [ No CAS ]
5-phenyl-N-[trans-3-[3-(1-hydroxyethyl)-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
57%	With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 20℃; for 2h;	27.3 [0357] Step 3: 5-phenyl-N-[trans-3- 13-(1-hydroxyethyl)-1H-pyrazol-1-ylj cyclobutyll - 1,2-oxazole-3-carboxamide: into a 50-mL round-bottom flask, was placed a solution of 5- phenyl-1,2-oxazole-3-carboxylic acid (177.7 mg, 0.94 mmol, 1.00 eq.), 1-[1-[trans-3- aminocyclobutyl]- 1 H-pyrazol-3-yl]ethan- 1 -ol hydrochloride (246 mg, 1.13 mmol, 1.20 eq.),HATU (428.8 mg, 1.13 mmol, 1.20 eq.) and DIEA (363.9 mg, 2.82 mmol, 3.00 eq.) in DMF(10 mL). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of 20 mL of water. The resulting solution was extracted with ethyl acetate (3x20 mL) and the organic combined layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a Prep-TLC with ethylacetate/petroleum ether (2:1). This resulted in 189 mg (57%) of 5 -phenyl-N-[trans-3 -[3 -(1- hydroxyethyl)- 1 H-pyrazol- 1 -yl]cyclobutyl] -1 ,2-oxazole-3 -carboxamide as a off-white solid.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0357

37
[ 14441-90-8 ]
[1-[cis-3-aminocyclobutyl]-1H-pyrazol-3-yl]methanol hydrochloride [ No CAS ]
[1-[cis-3-(5-phenyl-1,2-oxazole-3-amido)cyclobutyl]-1H-pyrazol-3-yl]methyl 5-phenyl-1,2-oxazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
40%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 2h;	30.4 [0383] Step 4: j1-jcis-3-(5-phenyl-1,2-oxazole-3-amido)cyclobutylj -1H-pyrazol-3- ylj methyl 5-phenyl-1,2-oxazole-3-carboxylate: into a 1 00-mL round-bottom flask, wasplaced a solution of [1-[cis-3-aminocyclobutyl]-1H-pyrazol-3-yl]methanol hydrochloride (400 mg, 1.96 mmol, 1.00 eq.), 5-phenyl-1,2-oxazole-3-carboxylic acid (745 mg, 3.94 mmol, 2.00 eq.), HCTU (983 mg, 2.36 mmol, 1.20 eq.) and DIEA (762 mg, 5.90 mmol, 3.00 eq.) in DMF (20 mL). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with ethyl acetateand the organic layers combined. The resulting mixture was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The crude residue was washed with 5 mL of methanol. This resulted in 400 mg (40%) of[1-[cis-3-(5-phenyl-1,2-oxazole-3- amido)cyclobutyl] -1 H-pyrazol-3 -yl]methyl 5 -phenyl- 1 ,2-oxazole-3 -carboxylate as a white solid. LC-MS: (M+H7 = 510.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0383

38
[ 14441-90-8 ]
[5-(3-aminocyclobutyl)-1,2,4-oxadiazol-3-yl]methanol [ No CAS ]
N-[3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]cyclobutyl]-5-phenyl-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1 g	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	32.7 [0399] Step 7: N-j3- 13-(hydroxymethyl)-1,2,4-oxadiazol-5-ylj cyclobutylj-5-phenyl-1,2- oxazole-3-carboxamide: into a 250-mL round-bottom flask, was placed a solution of 5-phenyl-1,2-oxazole-3-carboxylic acid (1 g, 5.28 mmol, 1.10 eq.) in dichloromethane (100 mL). To the mixture were added [5-(3-aminocyclobutyl)-1,2,4-oxadiazol-3-yl]methanol (800 mg, 4.52 mmol, 1.00 eq.) and HATU (2.16 g, 8.96 mmol, 1.20 eq.). This was followed by the addition of DIEA (2 g, 14.5 mmol, 3.00 eq.) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition ofwater. The resulting solution was extracted with dichloromethane (3 xl 00 mL) and the organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with dichloromethane/methanol (50:1). This resulted in 1g (62%) of N- [3 -[3 -(hydroxymethyl)- 1 ,2,4-oxadiazol-5-yl]cyclobutyl] -5-phenyl- 1 ,2-oxazole-3 -+carboxamide as a white solid. LC-MS: [M+H] = 341.
1 g	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	32.7 N-[3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]cyclobutyl]-5-phenyl-1,2-oxazole-3-carboxamide into a 250-mL round-bottom flask, was placed a solution of 5-phenyl- l,2-oxazole-3-carboxylic acid (1 g, 5.28 mmol, 1.10 eq.) in dichloromethane (100 mL). To the mixture were added [5-(3-aminocyclobutyl)-l,2,4-oxadiazol-3-yl]methanol (800 mg, 4.52 mmol, 1.00 eq.) and HATU (2.16 g, 8.96 mmol, 1.20 eq.). This was followed by the addition of DIEA (2 g, 14.5 mmol, 3.00 eq.) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with dichloromethane (3x100 mL) and the organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with dichloromethane/methanol (50: 1). This resulted in 1 g (62%) of N-[3-[3-(hydroxymethyl)-l,2,4-oxadiazol-5-yl]cyclobut l]-5-phenyl-l,2-oxazole-3- carboxamide as a white solid. LC-MS: [M+H]+ = 341

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0399
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0351

39
[ 14441-90-8 ]
[3-(3-aminocyclobutyl)-1,2,4-oxadiazol-5-yl]methyl acetate [ No CAS ]
[3-[3-(3-phenyl-1,2-oxazole-5-amido)cyclobutyl]-1,2,4-oxadiazol-5-yl]methyl acetate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 25℃; for 2h;	33.7 [0413] Step 7: 13-13-(3-phenyl-1,2-oxazole-5-amido)cyclobutylj -1,2,4-oxadiazol-5-ylimethyl acetate: into a 100-mL round-bottom flask, was placed a solution of [3-(3- aminocyclobutyl)- 1 ,2,4-oxadiazol-5-yl]methyl acetate (1.1 g, 5.21 mmol, 1.00 eq.) in dichloromethane (50 mL). To the solution were added DIEA (2.02 g, 15.63 mmol, 3.00 eq.), HCTU (3.25 g, 7.80 mmol, 1.50 eq.) and 3-phenyl-1,2-oxazole-5-carboxylic acid (1.18 g, 6.24mmol, 1.20 eq.). The resulting solution was stirred for 2 hours at 25 °C. The resulting solution was diluted with 150 mL of H20, extracted with ethyl acetate (2x150 mL) and the organic layers combined. The resulting mixture was washed with brine (2x200 mL), dried over anhydrous sodium sulfate and concentrated under vacuum to give 1.5 g (crude) of[3-[3-(3- phenyl- 1 ,2-oxazole-5-amido)cyclobutyl] -1 ,2,4-oxadiazol-5 -yl]methyl acetate as yellow oil.LC-MS [M+H] = 383.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0413

40
[ 14441-90-8 ]
(1S)-1-[5-[(3-aminocyclobutyl)methyl]-1,3,4-thiadiazol-2-yl]ethan-1-ol hydrochloride [ No CAS ]
(1S)-1-(5-[[3-(5-phenyl-1,2-oxazole-3-carboxamido)cyclobutyl]methyl]-1,3,4-thiadiazol-2-yl)ethyl 5-phenyl-1,2-oxazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
79%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 0 - 20℃; for 4h;	34.6 [0428] Step 6: (1 S)-1-(5- j 13-(5-phenyl-1,2-oxazole-3-amido)cyclobutylj methylj-1,3,4- thiadiazol-2-yl)ethyl 5-phenyl-1,2-oxazole-3-carboxylate: a solution of (1 S)- 1- [5- [(3- aminocyclobutyl)methyl] -1,3 ,4-thiadiazol-2-yl] ethan- 1 -ol hydrochloride (1.2 g, 4.80 mmol,1.00 eq.), 5-phenyl-1,2-oxazole-3-carboxylic acid (2.36 g, 12.48 mmol, 2.60 eq.) and HCTU (6.0 g, 14.50 mmol, 3.00 eq.) in dichloromethane (50 mL) was placed in a 100-mL round- bottom flask. This was followed by the addition of DIEA (3.1 g, 23.99 mmol, 5.00 eq.) dropwise with stirring at 0°C. The resulting solution was stirred for 4 hours at room temperature. The reaction was then quenched by the addition of 50 mL of water/ice andextracted with dichloromethane (3x50 mL) and the organic layers combined. The resulting mixture was washed with brine (3x30 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (2:1). This resulted in 2.1 g (79%) of(1S)-1-(5-[[3-(5-phenyl-1,2- oxazole-3 -amido)cyclobutyl]methyl]- 1,3 ,4-thiadiazol-2-yl)ethyl 5-phenyl- 1 ,2-oxazole-3 -carboxylate as a off-white solid. LC-MS: (M+H) = 556.
79%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 0 - 20℃;	34.6 Step 6: (lS)-l-(5-[[3-(5-phenyl-l,2-oxazole-3-amido)cyclobutyl]methyl]-l,3,4- thiadiazol-2-yl)ethyl 5-phenyl-l,2-oxazole-3-carboxylate: a solution of (lS)-l-[5-[(3- aminocyclobutyl)methyl]-l,3,4-thiadiazol-2-yl]ethan-l-ol hydrochloride (1.2 g, 4.80 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (2.36 g, 12.48 mmol, 2.60 eq.) and HCTU (6.0 g, 14.50 mmol, 3.00 eq.) in dichloromethane (50 mL) was placed in a 100-mL round- bottom flask. This was followed by the addition of DIEA (3.1 g, 23.99 mmol, 5.00 eq.) dropwise with stirring at 0°C. The resulting solution was stirred for 4 hours at room temperature. The reaction was then quenched by the addition of 50 mL of water/ice and extracted with dichloromethane (3x50 mL) and the organic layers combined. The resulting mixture was washed with brine (3x30 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (2: 1). This resulted in 2.1 g (79%) of (lS)-l-(5-[[3-(5-phenyl-l,2- oxazole-3-amido)cyclobutyl]methyl]-l,3,4-thiadiazol-2-yl)ethyl 5-phenyl-l,2-oxazole-3- carboxylate as a off-white solid. LC-MS: (M+H)+ = 556.
2.1 g	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 0 - 20℃; for 4h;	34.6 (1S)-1-(5-[[3-(5-phenyl-1,2-oxazole-3-amido)cyclobutyl]methyl]-1,3,4-thiadiazol-2-yl)ethyl 5-phenyl-1,2-oxazole-3-carboxylate a solution of (lS)-l-[5-[(3- aminocyclobutyl)methyl]-l,3,4-thiadiazol-2-yl]ethan-l-ol hydrochloride (1.2 g, 4.80 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (2.36 g, 12.48 mmol, 2.60 eq.) and HCTU (6.0 g, 14.50 mmol, 3.00 eq.) in dichloromethane (50 mL) was placed in a 100-mL round- bottom flask. This was followed by the addition of DIEA (3.1 g, 23.99 mmol, 5.00 eq.) dropwise with stirring at 0°C. The resulting solution was stirred for 4 hours at room temperature. The reaction was then quenched by the addition of 50 mL of water/ice and extracted with dichloromethane (3x50 mL) and the organic layers combined. The resulting mixture was washed with brine (3x30 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (2: 1). This resulted in 2.1 g (79%) of (lS)-l-(5-[[3-(5-phenyl-l,2- oxazole-3-amido)cyclobutyl]methyl]-l,3,4-thiadiazol-2-yl)ethyl 5-phenyl-l,2-oxazole-3- carboxylate as a off-white solid. LC-MS: (M+H)+ = 556

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0428
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0422
[3]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0378

41
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2-(3-aminocyclobutyl)ethan-1-ol hydrochloride [ No CAS ]
N-[3-(2-hydroxyethyl)cyclobutyl]-5-phenyl-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	36.2 Step 2: N-13-(2-hydroxyethyl)cyclobutylj -5-phenyl-1,2-oxazole-3-carboxamide: [0444] Step 2: N-13-(2-hydroxyethyl)cyclobutylj -5-phenyl-1,2-oxazole-3-carboxamide:a solution of 5-phenyl-1,2-oxazole-3-carboxylic acid (850.5 mg, 4.50 mmol, 1.51 eq.) and 2-(3- aminocyclobutyl)ethan-1-ol hydrochloride (452 mg, 2.98 mmol, 1.00 eq.) in dichloromethane(25 mL)was placed in a 100-mL round-bottom flask. HATU (1.368 g, 3.60 mmol, 1.21 eq.) and DIEA (1.161 g, 8.98 mmol, 3.01 eq.) were added to the solution and stirred for 1 hour at room temperature. The resulting solution was diluted with 50 mL of water, extracted with chloromethane (3x30 mL) and the organic layers combined. The resulting mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product waspurified by Flash-Prep-HPLC with the following conditions (CombiFlash-1): Column, C18 silica gel; mobile phase, MeCN/H20=55:45 increasing to MeCN/H20=60:40 within 2 mm; Detector, UV 254 nm to give 110 mg (13%) of N-[3-(2-hydroxyethyl)cyclobutyl]-5-phenyl- 1,2-oxazole-3-carboxamide as a off-white solid.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0444

42
[ 14441-90-8 ]
cis-3-aminocyclobutanecarbonitrile hydrochloride [ No CAS ]
N-(cis-3-cyanocyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
49%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	37.3 [0454] Step 3: N-(cis-3-cyanocyclobutyl)-5-phenylisoxazole-3-carboxamide: 3-aminocyclobutanecarbonitrile hydrochloride (440 mg, 4.58 mmol, 1.00 eq.), 5-phenyl-1,2-oxazole-3-carboxylic acid (866 mg, 4.58 mmol, 1.00 eq.) and HATU (2090 mg, 5.50 mmol, 1.20 eq.) in dichloromethane (18 mL) were placed in a 100-mL round-bottom flask. To the mixture was added DIEA (1773 mg, 13.72 mmol, 3.00 eq.) and the mixture was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. Theresulting solution was extracted with ethyl acetate and the organic layers combined. The resulting mixture was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:3) to give 600 mg (49%) of N-(cis-3-cyanocyclobutyl)-5- phenylisoxazole-3-carboxamide as a white solid. LC-MS: (M+H) = 268.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0454

43
[ 14441-90-8 ]
[3-([5-[(1R)-1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,3,4-oxadiazol-2-yl]methyl)cyclobutyl]amino 2,2,2-trffluoroacetate [ No CAS ]
N-[3-([5-[(1R)-1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,3,4-oxadiazol-2-yl]methyl)cyclobutyl]-5-phenyl-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
2.7 g	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 3h;	39.3 [0484] Step 3: N-j3-( 15- j(1R)-1- j(tert-butyldimethylsilyl)oxyj ethylj -1,3,4-oxadiazol-2- ylj methyl)cyclobutylj -5-phenyl-1,2-oxazole-3-carboxamide: a solution of [3 -([5 -[(1 R)- 1-[(tert-butyldimethylsilyl)oxy]ethyl] -1,3 ,4-oxadiazol-2-yl]methyl)cyclobutyl] amino 2,2,2-trifluoroacetate (3 g, 7.08 mmol, 1.00 eq.), 5-phenyl-1,2-oxazole-3-carboxylic acid (2.68 g,14.17 mmol, 2.00 eq.), HCTU (7.3 g, 17.65 mmol, 2.50 eq.) and DIEA (4.6 g, 35.59 mmol,5.00 eq.) in dichloromethane (100 mL) was stirred for 3 hours at room temperature. Theresulting solution was diluted with 100 mL of water, extracted with dichloromethane (3x100mL) and the organic layers combined. The resulting mixture was washed with brine (3x50mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:1) to give 2.7 g (79%) of N-[3 -([5 -[(1 R)- 1 -[(tert-butyldimethylsilyl)oxy] ethyl] -1,3 ,4-oxadiazol-2-yl]methyl)cyclobutyl]- 5-phenyl-1,2-oxazole-3-carboxamide as a white solid. LC-MS: (M+H7 = 483.
2.7 g	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 3h;	39.3 Step 3: ^-[3-([5-[(lR)-l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,3,4-oxadiazol-2- yl]methyl)cyclobutyl]-5-phenyl-l,2-oxazole-3-carboxamide: a solution of [3-([5-[(lR)-l- [(tert-bufyldimethylsilyl)oxy]ethyl]-l,3,4-oxadiazol-2-yl]methyl)cyclobufyl]amino 2,2,2- trifluoroacetate (3 g, 7.08 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (2.68 g, 14.17 mmol, 2.00 eq.), HCTU (7.3 g, 17.65 mmol, 2.50 eq.) and DIEA (4.6 g, 35.59 mmol, 5.00 eq.) in dichloromethane (100 mL) was stirred for 3 hours at room temperature. The resulting solution was diluted with 100 mL of water, extracted with dichloromethane (3x100 mL) and the organic layers combined. The resulting mixture was washed with brine (3x50 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 : 1) to give 2.7 g (79%) of N-[3-([5-[(lR)-l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,3,4-oxadiazol-2-yl]methyl)cyclobutyl]- 5-phenyl-l,2-oxazole-3-carboxamide as a white solid. LC-MS: (M+H)+ = 483.
2.7 g	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 3h;	39.3 N-[3-([5-[(1R)-1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,3,4-oxadiazol-2-yl]methyl)cyclobutyl]-5-phenyl-1,2-oxazole-3-carboxamide a solution of [3-([5-[(lR)-l- [(tert-butyldimethylsilyl)oxy]ethyl]-l,3,4-oxadiazol-2-yl]methyl)cyclobutyl]amino 2,2,2- trifluoroacetate (3 g, 7.08 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (2.68 g, 14.17 mmol, 2.00 eq.), HCTU (7.3 g, 17.65 mmol, 2.50 eq.) and DIEA (4.6 g, 35.59 mmol, 5.00 eq.) in dichloromethane (100 mL) was stirred for 3 hours at room temperature. The resulting solution was diluted with 100 mL of water, extracted with dichloromethane (3x100 mL) and the organic layers combined. The resulting mixture was washed with brine (3x50 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 : 1) to give 2.7 g (79%) of N-[3-([5-[(lR)-l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,3,4-oxadiazol-2-yl]methyl)cyclobutyl]- 5-phenyl-l,2-oxazole-3-carboxamide as a white solid. LC-MS: (M+H)+ = 483

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0484
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0477
[3]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0431

44
[ 14441-90-8 ]
(1R)-1-[3-(3-aminocyclobutyl)-1,2,4-oxadiazol-5-yl]ethan-1-ol [ No CAS ]
(1R)-1-[3-[3-(5-phenyl-1,2-oxazole-3-carboxamido)cyclobutyl]-1,2,4-oxadiazol-5-yl]ethyl 5-phenyl-1,2-oxazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
87%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 0 - 20℃; for 0.5h;	41.7 [0508] Step 7: (1R)-1- j3- 13-(5-phenyl-1,2-oxazole-3-amido)cyclobutylj -1,2,4-oxadiazol- 5-yllethyl 5-phenyl-1,2-oxazole-3-carboxylate: DIEA (1.95 g, 15.09 mmol, 5.00 eq.) wasadded dropwise to a 0°C solution of (1 R)- 1- [3 -(3 -aminocyclobutyl)- 1 ,2,4-oxadiazol-5 -yl] ethan1-ol (520 mg, 2.84 mmol, 1.00 eq.), 5-phenyl-1,2-oxazole-3-carboxylic acid (1.14 g, 6.03 mmol, 2.00 eq.) and HCTU (3.1 g, 7.49 mmol, 2.50 eq.) in dichloromethane (60 mL). The resulting solution was stirred for 30 mm at room temperature. The resulting solution was diluted with 50 mL of water/ice and extracted with dichloromethane (3x30 mL) and the organiclayers combined. The resulting mixture was washed with brine (2x20 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:1) to give 1.3 g (87%) of(1R)-1-[3-[3-(5-phenyl- 1 ,2-oxazole-3 -amido)cyclobutyl] -1 ,2,4-oxadiazol-5-yl] ethyl 5-phenyl- 1 ,2-oxazole-3 -+carboxylate as light yellow oil. LC-MS: (M+H) = 526.
87%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 0 - 20℃;	41.7 Step 7: (lR)-l-[3-[3-(5-phenyl-l,2-oxazole-3-amido)cyclobutyl]-l,2,4-oxadiazol- 5-yl]ethyl 5-phenyl-l,2-oxazole-3-carboxylate: DIEA (1.95 g, 15.09 mmol, 5.00 eq.) was added dropwise to a 0°C solution of (lR)-l-[3-(3-aminocyclobutyl)-l,2,4-oxadiazol-5-yl]ethan- l-ol (520 mg, 2.84 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (1.14 g, 6.03 mmol, 2.00 eq.) and HCTU (3.1 g, 7.49 mmol, 2.50 eq.) in dichloromethane (60 mL). The resulting solution was stirred for 30 min at room temperature. The resulting solution was diluted with 50 mL of water/ice and extracted with dichloromethane (3x30 mL) and the organic layers combined. The resulting mixture was washed with brine (2x20 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 : 1) to give 1.3 g (87%) of (lR)-l-[3-[3- (5-phenyl-l,2-oxazole-3-amido)cyclobutyl]-l,2,4-oxadiazol-5-yl] ethyl 5-phenyl-l,2-oxazole-3- carboxylate as light yellow oil. LC-MS: (M+H)+ = 526
87%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 0 - 20℃; for 0.5h;	41.7 (1R)-1-[3-[3-(5-phenyl-1,2-oxazole-3-amido)cyclobutyl]-1,2,4-oxadiazol-5-yl]ethyl 5-phenyl-1,2-oxazole-3-carboxylate DIEA (1.95 g, 15.09 mmol, 5.00 eq.) was added dropwise to a 0°C solution of (lR)-l-[3-(3-aminocyclobutyl)-l,2,4-oxadiazol-5-yl]ethan- l-ol (520 mg, 2.84 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (1.14 g, 6.03 mmol, 2.00 eq.) and HCTU (3.1 g, 7.49 mmol, 2.50 eq.) in dichloromethane (60 mL). The resulting solution was stirred for 30 min at room temperature. The resulting solution was diluted with 50 mL of water/ice and extracted with dichloromethane (3x30 mL) and the organic layers combined. The resulting mixture was washed with brine (2x20 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 : 1) to give 1.3 g (87%) of (lR)-l-[3-[3- (5-phenyl-l,2-oxazole-3-amido)cyclobu†yl]-l,2,4-oxadiazol-5-yl] ethyl 5-phenyl-l,2-oxazole-3- carboxylate as light yellow oil. LC-MS: (M+H)+ = 526.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0508
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0499
[3]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0453

45
[ 14441-90-8 ]
trans-3-[5-[(1R)-1-methoxyethyl]-1,3,4-oxadiazol-2-yl]cyclobutan-1-amine [ No CAS ]
5-phenyl-N-[trans-3-[5-[(1R)-1-methoxyethyl]-1,3,4-oxadiazol-2-yl]cyclobutyl]isoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
70%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 0 - 20℃; for 1h;	45.6 [0534] Step 6: 5-phenyl-N- jtrans-3 - j5- j(1R)-1-methoxyethylj-1,3,4-oxadiazol-2- yljcyclobutyljisoxazole-3-carboxamide: DIEA (943 mg, 7.30 mmol, 3.00 eq.) was added dropwise to a cold solution of 5-phenylisoxazole-3-carboxylic acid (550 mg, 2.91 mmol, 1.20 eq.), trans-3 -[5- [(1 R)- 1 -methoxyethyl] -1,3 ,4-oxadiazol-2-yl]cyclobutan- 1-amine (480 mg, 2.43mmol, 1.00 eq.) and HATU (1.387 g, 3.65 mmol, 1.50 eq.) in dichloromethane (50 mL) at 0 °C. The resulting solution was stirred for 1 hour at room temperature and then diluted with 50 mL of dichloromethane. The resulting mixture was washed with water (2x50 mL) and brine (1x50 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:1) to give 628 mg (70%)of 5 -phenyl-N- [trans-3 -[5- [(1 R)- 1 -methoxyethyl]- 1,3 ,4-oxadiazol-2-yl]cyclobutyl] isoxazole-3 - carboxamide as an off-white solid.[0535] Analytical data:[0536] HPLC purity: 98.9% at 254 nm[0537] LC-MS (ES, m/z): [M+1] = 369[0538] ‘H NMR (400MHz, DM50-cl6): 9.33-9.31 (d, J= 7.6 Hz, 1H), 7.96-7.94 (t, J= 5.6Hz, 2H), 7.59-7.56 (m, 3H), 7.38 (s, 1H), 4.74-4.67 (m, 2H), 3.76-3.70 (m, 1H),3.29 (s, 3H), 2.74-2.61 (m, 4H), 1.5 1-1.49 (d, J= 6.8 Hz, 3H).

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0534; 0535; 0536; 0537; 0538

46
[ 14441-90-8 ]
trans-3-(1H-imidazol-1-yl)cyclobutan-1-amine hydrochloride [ No CAS ]
5-phenyl-N-[trans-3-(1H-imidazol-1-yl)cyclobutyl]isoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
6%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	62.4 [0629] Step 4: 5-phenyl-N- jtrans-3-(1H-imidazol-1-yl)cyclobutylj isoxazole-3- carboxamide: trans-3 -(1 H-imidazol- 1 -yl)cyclobutan- 1-amine hydrochloride (400 mg, 2.30mmol, 0.70 eq.), HATU (1.507 g, 3.96 mmol, 1.20 eq.) and DIEA (1.279 g, 9.90 mmol, 3.00 eq.) were added to a solution of 5-phenylisoxazole-3-carboxylic acid (624 mg, 3.30 mmol, 1.00 eq.) in dichloromethane (30 mL). The resulting solution was stirred for 2 hours at room temperature and it was then concentrated under vacuum. The crude product was purified by Prep-HPLC with the following conditions (Waters): Column, XBridge BEH 130 Prep C18 OBDColumn, 19*150mm 5um l3nm; mobile phase, water with 0.05%TFA and ACN (12.0% ACN up to 50.0% in 8 mm); Detector, UV 254nm. This resulted in 58 mg (6%) of 5-phenyl-N- [trans-3 -(1 H-imidazol- 1 -yl)cyclobutyl]isoxazole-3 -carboxamide as a white solid.[0630] Analytical data:[0631] LC-MS (ES, m/z): [M+H] = 309.2[0632] HPLC purity: 99.4% at 254 nm[0633] ‘H NMR (300MHz, DMSO-d6,ppm): ö 9.28 (s, 1H), 7.98-7.93 (m, 3H), 7.51 (s, 1H), 7.60-7.56 (m, 3H), 7.37 (s, 1H), 5.20-5.11 (m, 1H), 4.69-4.60 (m, 1H),2.91-2.45 (m, 4H).

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0629; 0630; 0631; 0632; 0633

47
[ 14441-90-8 ]
(1R)-1-(5-(3-aminocyclohexyl)-1,3,4-oxadiazol-2-yl)ethan-1-ol [ No CAS ]
(1R)-1-[5-[3-(5-phenylisoxazole-3-amido)cyclohexyl]-1,3,4-oxadiazol-2-yl]ethyl 5-phenylisoxazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
350 mg	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	63.5 [0638] Step 5: (1R)-1- j5- 13-(5-phenylisoxazole-3-amido)cyclohexylj -1,3,4-oxadiazol-2-yllethyl 5-phenylisoxazole-3-carboxylate 5-phenylisoxazole-3-carboxylic acid (275 mg, 1.45mmol, 1.50 eq.), HATU (553 mg, 1.45 mmol, 1.50 eq.) and DIEA (376 mg, 2.91 mmol, 3.00eq.) were added to a solution of (1 R)- 1 -(5 -(3 -aminocyclohexyl)- 1,3 ,4-oxadiazol-2-yl)ethan- 1 -ol(300 mg, 0.97 mmol, 1.00 eq.) in dichloromethane (20 mL). The resulting solution was stirred for 2 hours at room temperature and it was then concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:5) to give 350 mg (65%) of (1 R)- 1- [5- [3 -(5 -phenylisoxazole-3 -amido)cyclohexyl] -1,3 ,4-oxadiazol-2-yl]ethyl 5-phenylisoxazole-3-carboxylate as a white solid. LC-MS (ES, m/z): [M+H] = 554.3.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0638

48
[ 14441-90-8 ]
trans-3-(1H-1,3-benzodiazol-1-yl)cyclobutan-1-amine [ No CAS ]
5-phenyl-N-[trans-3-(1H-1,3-benzodiazol-1-yl)cyclobutyl]isoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
37.2 mg	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃;	67.7 [0689] Step 7: 5-phenyl-N- jtrans-3-(1H-1,3-benzodiazol-1-yl)cyclobutylj isoxazole-3- carboxamide: 5-phenylisoxazole-3-carboxylic acid (374 mg, 1 .98mmol, 1.00 eq.), HATU (902 mg, 2.37mmol, 1.20 eq.) and DIEA (766 mg, 5.93mmol, 3.00 eq.) were added to solution oftrans-3-(1H-1,3-benzodiazol-1-yl)cyclobutan-1-amine (370 mg, 1.98mmol, 1.00 eq.) in dichloromethane (5mL) and the solution was stirred for overnight at room temperature. The reaction was then quenched by the addition of lOmL of water, extracted with dichloromethane (3xl5mL) and the organic layers combined. The resulting mixture was washed with brine (3x25mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The crudeproduct was purified by Prep-HPLC with the following conditions (HPLC-10): Column, X Bridge BEH13O Prep C18 OBD Column, l9xlSOmm, Sum l3nm; mobile phase, Water with 0.05% NH4HCO3 and ACN (37% ACN up to 52% in 8 mm); Detector, UV 254 nm. This resulted in 37.2 mg (5%) of 5-phenyl-N-[trans-3-(1H-1,3-benzodiazol-1- yl)cyclobutyl]isoxazole-3-carboxamide as a light yellow solid.[0690] Analytical data:[0691] LC-MS (ES, m/z): [M+H] = 359.2[0692] HPLC purity: 98.9% at 254 nm[0693] ‘H NMR (CDC13, 400 MHz): ö 8.39 (s, 1H), 7.89-7.81 (m, 3H), 7.52-7.50 (m, 3H), 7.48-7.43 (m, 1H), 7.39-7.35 (m, 2H), 7.00 (s, 1H), 5.25-5.2 1 (m, 1H), 4.8 1-4.79(br, 1H), 3.17-3.10 (m, 2H), 2.98-2.95 (m, 2H).

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0689; 0690; 0691; 0692; 0693

49
[ 14441-90-8 ]
[1-[cis-3-aminocyclobutyl]-1H-pyrazol-4-yl]methanol hydrochloride [ No CAS ]
5-phenyl-N-[cis-3-[4-(hydroxymethyl)-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 20℃; for 1h;	28.3 [0372] Step 3: 5-phenyl-N- jcis-3- 14-(hydroxymethyl)-1H-pyrazol-1-ylj cyclobutylj -1,2- oxazole-3-carboxamide: into a 50-mL round-bottom flask, was placed a solution of [1-[cis-3- aminocyclobutyl]-1H-pyrazol-4-yl]methanol hydrochloride (408 mg, 2.00 mmol, 1.20 eq.) in DMF (10 mL). To the solution were added DIEA (645 mg, 4.99 mmol, 3.00 eq.), 5-phenyl-1,2-oxazole-3-carboxylic acid (315 mg, 1.67 mmol, 1.00 eq.) and HATU (760 mg, 2.00 mmol,1.20 eq.). The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of 20 mL of water. The resulting solution was extracted with ethyl acetate (3x20 mL) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a Prep-TLC with ethylacetate/petroleum ether (2:1). This resulted in 63 mg (11%) of 5-phenyl-N-[cis-3-[4- (hydroxymethyl)- 1 H-pyrazol- 1 -yl]cyclobutyl]- 1 ,2-oxazole-3 -carboxamide. [0373] Appearance: white solid[0374] Analytical data: ‘HNMR (400MHz, DMSO-d6): ö 9.24, 9.22 (d, J= 8.0Hz, 1H),7.95-7.92 (m, 2H), 7.80 (s, 1H), 7.59-7.53 (m, 3H), 7.38, 7.36 (d, J= 8.0Hz,2H), 4.83-4.80 (t, J= 6.0Hz, 1H), 4.60-4.56 (m, 1H), 4.36-4.28 (m, 3H), 2.82-2.75 (m, 2H), 2.65-2.59 (m, 2H).[0375] LC-MS: (M+H) = 339[0376] HPLC purity: 99.94% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0372; 0373; 0374; 0375; 0376

50
[ 14441-90-8 ]
[1-[trans-3-aminocyclobutyl]-1H-pyrazol-3-yl]methanol [ No CAS ]
N-(trans-3-(3-(hydroxymethyl)-1H-pyrazol-1-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
18%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 1h;	31 Example 31: N-trans-3-(3-(hydroxymethyl)-1H-pyrazol-1-yl)cyclobutyl)-5- phenylisoxazole-3-carboxamide [0388] Into a 100-mL round-bottom flask, was placed a solution of [1-trans-3- aminocyclobutyl]-1H-pyrazol-3-yl]methanol (120 mg, 0.72 mmol, 1.00 eq., prepared usingsimilar procedure as shown in example 29) in dichloromethane (5 mL). To the solution were added 5-phenyl-1,2-oxazole-3-carboxylic acid (163 mg, 0.86 mmol, 1.20 eq.) and HCTU (360 mg, 0.87 mmol, 1.20 eq.). This was followed by the addition of DIEA (278 mg, 2.15 mmol, 3.00 eq.) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water. The resulting solutionwas extracted with dichloromethane (3x50 mL). The organic layers were combined, dried and concentrated under vacuum. The crude product was purified by Prep-HPLC with the following conditions (Waters): Column, Bridget Prep C18 Sum OBDTM 19*100mm; mobile phase, water with 0.05% NH4HCO3 and CH3CN (40.0% CH3CN up to 80.0% in 10 mm, up to 95.0% in 1.5mm, down to 40.0% in 1.5mm); Detector, 254nm. This resulted in 44.7 mg (18%) of 5-phenyl-N-trans-3 -[3 -(hydroxymethyl)- 1 H-pyrazol- 1 -yl]cyclobutyl] -1 ,2-oxazole-3 -carboxamide as a white solid.[0389] LC-MS: (M+H) = 339[0390] Analytical data: ‘H NMR (400MHz, DMSO-d6): ö 9.32-9.30 (d, J= 6.8 Hz, 1H),7.95-7.94 (d, J= 6.0 Hz, 2H), 7.74 (s, 1H), 7.57-7.55 (m, 3H), 7.38 (s, 1H), 6.20(s, 1H), 5.02-4.99 (t, J= 5.6Hz, 1H), 4.96-4.95 (m, 1H), 4.7 1-4.65 (m, 1H), 4.44-4.42 (d, J= 6.0 Hz, 2H), 2.75-2.63 (m, 4H).[0391] HPLC purity: 98.8% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0388; 0389; 0390; 0391

51
[ 14441-90-8 ]
(5-(3-(aminomethyl)cyclobutyl)-1,3,4-thiadiazol-2-yl)methanol hydrochloride [ No CAS ]
[5-(3-[[(5-phenyl-1,2-oxazol-3-yl)formamido]methyl]cyclobutyl)-1,3,4-thiadiazol-2-yl]methyl 5-phenyl-1,2-oxazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 25℃; for 3h;	4.4a [0183] Step 4a: (5-(3-((5-phenylisoxazole-3-carboxamido)methyl)cyclobutyl)-1,3,4- thiadiazol-2-yl)methyl 5-phenylisoxazole-3-carboxylate: a solution of [5-[3- (aminomethyl)cyclobutyl] -1,3 ,4-thiadiazol-2-yl]methanol hydrogen chloride (750 mg, 3.17 mmol, 1.00 eq.), 5-phenyl-1,2-oxazole-3-carboxylic acid (860 mg, 4.55 mmol, 1.40 eq.),HCTU (1.59 g, 3.82 mmol, 1.20 eq.) and DIEA (1.66 g, 12.84 mmol, 3.00 eq.) in dichloromethane (50 mL) was stirred for 3 hours at 25 °C. The resulting mixture was concentrated under vacuum. This resulted in 800 mg (crude) [5-(3-[[(5-phenyl-1,2-oxazol-3- yl)formamido]methyl]cyclobutyl)- 1,3 ,4-thiadiazol-2-yl]methyl 5-phenyl- 1 ,2-oxazole-3 - carboxylate as a yellow oil. The crude product was used in the next step directly without furtherpurification. LC-MS: 542.0 [M+H].
	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 25℃; for 3h;	4.4a Step 4a: (5-(3-((5-phenylisoxazole-3-carboxamido)methyl)cyclobutyl)-l,3,4- thiadiazol-2-yl)methyl 5-phenylisoxazole-3-carboxylate: a solution of [5-[3- (aminomethyl)cyclobutyl]-l,3,4-thiadiazol-2-yl]methanol hydrogen chloride (750 mg, 3.17 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (860 mg, 4.55 mmol, 1.40 eq.), HCTU (1.59 g, 3.82 mmol, 1.20 eq.) and DIEA (1.66 g, 12.84 mmol, 3.00 eq.) in dichloromethane (50 mL) was stirred for 3 hours at 25 °C. The resulting mixture was concentrated under vacuum. This resulted in 800 mg (crude) [5-(3-[[(5-phenyl-l,2-oxazol-3- yl)formamido]methyl]cyclobutyl)-l,3,4-thiadiazol-2-yl]methyl 5-phenyl-l,2-oxazole-3- carboxylate as a yellow oil. The crude product was used in the next step directly without further purification. LC-MS: 542.0 [M+H]+.
	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 25℃; for 3h;	4.4a [5-(3-[[(5-phenyl-1,2-oxazol-3-yl)formamido]methyl]cyclobutyl)-1,3,4-thiadiazol-2-yl]methyl 5-phenyl-1,2-oxazole-3-carboxylate a solution of [5-[3- (aminomethyl)cyclobutyl]-l,3,4-thiadiazol-2-yl]methanol hydrogen chloride (750 mg, 3.17 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (860 mg, 4.55 mmol, 1.40 eq.), HCTU (1.59 g, 3.82 mmol, 1.20 eq.) and DIEA (1.66 g, 12.84 mmol, 3.00 eq.) in dichloromethane (50 mL) was stirred for 3 hours at 25 °C. The resulting mixture was concentrated under vacuum. This resulted in 800 mg (crude) [5-(3-[[(5-phenyl-l,2-oxazol-3- yl)formamido]methyl]cyclobutyl)-l,3,4-thiadiazol-2-yl]methyl 5-phenyl-l,2-oxazole-3- carboxylate as a yellow oil. The crude product was used in the next step directly without further purification. LC-MS: 542.0 [M+H]+.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0183
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0180
[3]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0139

52
[ 14441-90-8 ]
3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutan-1-amine [ No CAS ]
5-phenyl-N-[cis-3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]
5-phenyl-N-[trans-3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 13% 2: 59%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	38.1 [0463] Step: 5-phenyl-N-jcis-3-(3- 11- j(tert-butyldimethylsilyl)oxyj ethylj -1,2,4-oxadiazol-5-yl)cyclobutylj-1,2-oxazole-3-carboxamide and 5-phenyl-N-jtrans-3-(3- 11-j(tert-butyldimethylsilyl)oxyj ethyll -1,2,4-oxadiazol-5-yl)cyclobutylj -1,2-oxazole-3-carboxamide: to a solution of 3 -(3 -[1- [(tert-butyldimethylsilyl)oxy]ethyl]- 1 ,2,4-oxadiazol-5 -yl)cyclobutan-1-amine (1.5 g, 5.04 mmol, 1.00 eq.) in dichloromethane (100 mL)was added 5-phenyl-1,2-oxazole-3-carboxylic acid (1.13 g, 5.97 mmol, 1.20 eq.), HATU (2.28 g, 6.00mmol, 1.20 eq.) and DIEA (1.93 g, 14.93 mmol, 3.00 eq.). The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water and extracted with ethyl acetate (3x50 mL) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto asilica gel column with ethyl acetate/petroleum ether (1 :50)to give 300 mg (13%) of 5-phenyl- N-[cis-3 -(3 -[1 -[(tert-butyldimethylsilyl)oxy] ethyl] -1 ,2,4-oxadiazol-5-yl)cyclobutyl]- 1,2- oxazole-3-carboxamide as a white solid. The solvent was changed to a mixture of with ethyl acetate/petroleum ether (1:20) to give 1.4 g (59%) of 5-phenyl-N-[trans-3 -(3 -[1 -[(tertbutyldimethylsilyl)oxy]ethyl]- 1 ,2,4-oxadiazol-5 -yl)cyclobutyl] -1 ,2-oxazole-3 -carboxamide as awhite solid. LC-MS: (M+H) = 469.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0463

53
[ 14441-90-8 ]
methyl 3-aminocyclopentane-1-carboxylate [ No CAS ]
methyl 3-(5-phenylisoxazole-3-carboxamido)cyclopentane-1-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
84%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	65.1 [0656] Step 1: methyl 3-(5-phenylisoxazole-3-carboxamido)cyclopentane-1- carboxylate: into a 50-mL round-bottom flask, was placed a solution of methyl 3- aminocyclopentane-1-carboxylate (500 mg, 3.49 mmol, 1.00 eq.) in dichloromethane (10 mL). To the solution were added HATU (1.59 g, 4.18 mmol, 1.20 eq.), DIEA (1.6 g, 12.38 mmol,3.50 eq.) and 5-phenylisoxazole-3-carboxylic acid (790 mg, 4.18 mmol, 1.20 eq.). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in 0.925 g (84%) of methyl 3-(5-phenylisoxazole-3-amido)cyclopentane- 1 -carboxylate as a light yellow solid. LC-MS (ES, m/z) [M+H] = 315.1
84%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	65.1 methyl 3-(5-phenylisoxazole-3-amido)cyclopentane-1-carboxylate into a 50-mL round-bottom flask, was placed a solution of methyl 3- aminocyclopentane-l-carboxylate (500 mg, 3.49 mmol, 1.00 eq.) in dichloromethane (10 mL). To the solution were added HATU (1.59 g, 4.18 mmol, 1.20 eq.), DIEA (1.6 g, 12.38 mmol, 3.50 eq.) and 5-phenylisoxazole-3-carboxylic acid (790 mg, 4.18 mmol, 1.20 eq.). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in 0.925 g (84%) of methyl 3-(5-phenylisoxazole-3- amido)cyclopentane-l-carboxylate as a light yellow solid. LC-MS (ES, m/z) [M+H]+ = 315.1

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0656
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0603

54
[ 14441-90-8 ]
ethyl trans-3-aminocyclobutane-1-carboxylate hydrochloride [ No CAS ]
ethyl trans-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-1-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
80%	With triethylamine; HATU In tetrahydrofuran at 20℃; for 6h;	9.6 [0239] Step 6: ethyl trans-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-1- carboxylate: Et3N (5.6 mL, 42 mmol) and HATU (4.84 g, 13 mmol) were added to a mixture of ethyl trans-3-aminocyclobutane-1-carboxylate hydrochloride (1.89 g, 10 mmcl) and 5-phenyliscxazcle-3-carbcxylic acid (2 g, 10 mmcl) in THF (200 mL) at room temperature and the reaction mixture was stirred for 6 h at room temperature. Volatiles were removed under reduced pressure to get the crude compound. The reaction mixture was diluted with water (100 mL) and extracted using ethyl acetate (2 x 75 mL). Combined organic layer was washed with brine (50 mL), dried over anhydrous Na2SO4 and concentrated under reduced pressure. Themixture was purified by flash column chromatography using 30% EtOAc in hexane as eluent to give the product (2.65 g, 80 %) as white solid. ‘H NMR (400 MHz, CDC13) : ö 7.80-7.77 (m, 2H), 7.49-7.46 (m, 3H), 7.00 (d, J= 7.2 Hz, 1H), 6.94 (s, 1H), 4.79-4.73 (m, 1H), 4.17 (q, J=7.1 Hz, 2H), 3.10-3.09 (m, 1H), 2.78-2.72 (m, 2H), 2.40-2.32 (m, 2H), 1.30-1.26 (t, J 7.2 Hz, 3H). LC-MS: [M+H] + 315.2

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0239

55
[ 14441-90-8 ]
(S)-3-amino-N'-(2-((tert-butyldimethylsilyl)oxy)propanoyl)cyclobutane-1-carbohydrazide [ No CAS ]
(S)-N-(3-(2-(2-((tert-butyldimethylsilyl)oxy)propanoyl)hydrazine-1-carbonyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
73.3%	With triethylamine; HATU In tetrahydrofuran at 20℃; for 3h;	18.1 Step 1: (S)-^-(3-(2-(2-((tert-butyldimethylsilyl)oxy)propanoyl)hydrazine-l- carbonyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: HATU (3.6 g, 9.5 mmol) was added to a solution of 5-phenylisoxazole-3-carboxylic acid (1.19 g, 6.3 mmol) in THF (20 mL) followed by addition of (S)-3-amino-N'-(2-((tert- butyldimethylsilyl)oxy)propanoyl)cyclobutane-l-carbohydrazide (2.0 g, 6.31 mmol). The reaction mixture was stirred for 10 minutes at room temperature and tri ethyl amine (2.67 mL, 19.0 mmol) was then added. The reaction mixture was stirred at room temperature for 3h, volatiles were removed under reduced pressure and the reaction mixture was quenched with ice-water (20 mL). The aq. phase was extracted with ethyl acetate (3 x 20 mL). Combined organic layer was washed with brine (20 mL), dried over Na2SC>4 and concentrated under reduced pressure to get the crude compound. The crude compound thus obtained was purified by combifiash using 45% ethyl acetate in hexane to get the product (2.2 g, 73.3 %) as a white solid. NMR (400 MHz, CDC13): δ 9.10 - 9.05 (m, 1H), 8.43 - 8.39 (m, 1H), 7.79 - 7.27 (m, 2H), 7.50 - 7.46 (m, 3H), 7.20 - 7.16 (m, 1H), 6.92 (s, 1H), 4.63 - 4.57 (m, 1H), 4.37 - 4.32 (m, 4H), 2.84 - 2.81 (m, 5H), 2.72 - 2.70 (m, 1H), 1.42 (d, J = 6.6 Hz, 2H), 0.95 (s, 9H), 0.14 (s, 3H), 0.12 (s, 3H); LC-MS: [M+H]+ 487.3.
73.3%	With triethylamine; HATU In tetrahydrofuran at 20℃; for 3h;	18.1 (S)-N-(3-(2-(2-((tert-butyldimethylsilyl)oxy)propanoyl)hydrazine-1-carbonyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide HATU (3.6 g, 9.5 mmol) was added to a solution of 5-phenylisoxazole-3-carboxylic acid (1.19 g, 6.3 mmol) in THF (20 mL) followed by addition of (S)-3-amino-N'-(2-((tert-butyldimethylsilyl)oxy)propanoyl) cyclobutane-l-carbohydrazide (2.0 g, 6.31 mmol). The reaction mixture was stirred for 10 minutes at room temperature and triethyl amine (2.67 mL, 19.0 mmol) was then added. The reaction mixture was stirred at room temperature for 3h, volatiles were removed under reduced pressure and the reaction mixture was quenched with ice-water (20 mL). The aq. phase was extracted with ethyl acetate (3 x 20 mL). Combined organic layer was washed with brine (20 mL), dried over Na2SC>4 and concentrated under reduced pressure to get the crude compound. The crude compound thus obtained was purified by combifiash using 45% ethyl acetate in hexane to get the product (2.2 g, 73.3 %) as a white solid. XH NMR (400 MHz, CDC13): δ 9.10 - 9.05 (m, 1H), 8.43 - 8.39 (m, 1H), 7.79 - 7.27 (m, 2H), 7.50 - 7.46 (m, 3H), 7.20 - 7.16 (m, 1H), 6.92 (s, 1H), 4.63 - 4.57 (m, 1H), 4.37 - 4.32 (m, 4H), 2.84 - 2.81 (m, 5H), 2.72 - 2.70 (m, 1H), 1.42 (d, J = 6.6 Hz, 2H), 0.95 (s, 9H), 0.14 (s, 3H), 0.12 (s, 3H); LC-MS: [M+H]+ 487.3.
2.2 g	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid; (S)-3-amino-N'-(2-((tert-butyldimethylsilyl)oxy)propanoyl)cyclobutane-1-carbohydrazide With HATU In tetrahydrofuran at 20℃; for 0.166667h; Stage #2: With triethylamine In tetrahydrofuran at 20℃; for 3h;	18.1 [0296] Step 1: (S)-N-(3-(2-(2-((tert-butyldimethylsilyl)oxy)propanoyl)hydrazine-1-carbonyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: HATU (3.6 g, 9.5 mmol) was added to a solution of 5-phenylisoxazole-3-carboxylic acid (1.19 g, 6.3 mmcl) in THF (20 mL) followed by addition of (S)-3-amino-N’-(2-((tert- butyldimethylsilyl)oxy)propanoyl)cyclobutane-1-carbohydrazide (2.0 g, 6.31 mmol). Thereaction mixture was stirred for 10 minutes at room temperature and triethyl amine (2.67 mL,19.0 mmol) was then added. The reaction mixture was stirred at room temperature for 3h, volatiles were removed under reduced pressure and the reaction mixture was quenched with ice-water (20 mL). The aq. phase was extracted with ethyl acetate (3 x 20 mL). Combined organic layer was washed with brine (20 mL), dried over Na2SO4 and concentrated underreduced pressure to get the crude compound. The crude compound thus obtained was purified by combiflash using 45% ethyl acetate in hexane to get the product (2.2 g, 73.3 %) as a white solid. ‘H NMR (400 MHz, CDC13): ö 9.10 - 9.05 (m, 1H), 8.43 - 8.39 (m, 1H), 7.79 - 7.27 (m, 2H), 7.50 - 7.46 (m, 3H), 7.20 - 7.16 (m, 1H), 6.92 (s, 1H), 4.63 -4.57 (m, 1H), 4.37 -4.32 (m, 4H), 2.84-2.81 (m, 5H), 2.72 -2.70 (m, 1H), 1.42 (d, J= 6.6 Hz, 2H), 0.95 (s, 9H), 0.14(s, 3H), 0.12 (s, 3H); LC-MS: [M+H] 487.3.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0290
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0249
[3]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0296

56
[ 14441-90-8 ]
(R)-1-(5-(3-aminocyclobutyl)-1,3,4-thiadiazol-2-yl)ethan-1-ol [ No CAS ]
N-((1S,3s)-3-(5-((R)-1-hydroxyethyl)-1,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
0.110 g	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid; (R)-1-(5-(3-aminocyclobutyl)-1,3,4-thiadiazol-2-yl)ethan-1-ol With HATU In tetrahydrofuran for 0.166667h; Stage #2: With triethylamine In tetrahydrofuran at 20℃; for 12h;	19.4; 19.5 [0305] Steps 4 and 5: N-((1S,3s)-3-(5-((R)-1-hydroxyethyl)-1,3,4-thiadiazol-2- yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: HATU (0.837 g, 2.2 mmol) was added to a solution of 5-phenylisoxazole-3-carboxylic acid (0.277 g, 1.4 mmol) in THF (5 mL) followedby addition of (R)- 1 -(5 -(3 -aminocyclobutyl)- 1,3 ,4-thiadiazol-2-yl)ethan- 1 -ol (0.46 g, crude) and the resulting reaction mixture was stirred for 10 mm. Triethyl amine (0.61 mL, 4.4 mmol) was added to the reaction mixture and stirring continued at room temperature for 12h. Cold water (20 mL) was added to the mixture and then extracted with DCM (2 x 10 mL). Combined organic layer was washed with brine, dried over Na2SO4 and evaporated to dryness undervacuum. The crude compound was dissolved in THF (5 mL) and TBAF solution (1.2 mL, 1.2 mmol) was added and the reaction mixture was stirred for lh. After completion, the reaction mixture was quenched with cold water (20 mL) and extracted with DCM (2 x 5 mL). Combined organic layer was washed with brine and dried over Na2SO4 and evaporated to dryness under reduced pressure to get the crude compound which was purified by prep HPLCto afford 5 -Phenyl-isoxazole-3 -carboxylic acid {3 -[5 -((R)- 1 -hydroxy-ethyl)- [1, 3 ,4]thiadiazol-2-yl]-cyclobutyl}-amide (0.110 g, 15 % over two steps) as an off white solid.[0306] Analytical data: ‘H NMR (400 MHz, CDC13): ö 7.78 - 7.76 (m, 2H), 7.50 - 7.46 (m,3H), 7. 20 (d, J= 8 Hz, 1H), 6.94 (s, 1H), 5.28 (q, J= 6.5 Hz, 1H), 4.71 - 4.65(m, 1H), 3.72 - 3.63 (m, 1H), 3.05 - 2.99 (m, 2H), 2.85 (br, 1H), 2.53 - 2.46 (m,2H), 1.68 (d, J= 6.5Hz, 3H).[0307] LC-MS: [M+H] 370.9[0308] HPLC purity: 98.2 1% at 220 nm and 98.95% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0305

57
[ 14441-90-8 ]
ethyl cis-3-aminocyclobutane-1-carboxylate hydrochloride [ No CAS ]
ethyl cis-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-1-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
79%	With triethylamine; HATU In tetrahydrofuran at 20℃; for 6h;	20.3 [0311] Step 3: ethyl cis-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-1-carboxylate: Et3N (5.3 mL, 0.04 mol) followed by HATU (9.16 g, 0.024 mol) were added to a solution of ethyl cis-3-aminocyclobutane-1-carboxylate hydrochloride (3.63 g, 0.020 mol) and 5-phenylisoxazole-3-carboxylic acid (4.20 g, 0.022 mol) in THF (150 mL)and the reaction mixture was stirred for 6 h at room temperature. Volatiles were removed under reducedpressure and the crude reaction mixture was diluted with water (100 mL). The aq. phase was extracted with ethyl acetate (200 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain the crude product. The crude compound was purified by silica gel column chromatography using 50% EtOAc in hexane as eluent to afford the product (5.02 g, 79 %) as off white solid. ‘H NMR (400 MHz, CDC13): ö 7.79 -7.77 (m,2H), 7.50 - 7.46 (m, 3H), 7.08 (d, J= 8.0 Hz, 1H), 6.93 (s, 1H), 4.61 -4.12 (m, 1H), 4.15 (q, J = 7.1 Hz, 2H), 2.89 -2.83 (m, 1H), 2.74 - 2.68 (m, 2H), 2.32 -2.24 (m, 2H), 1.26 (t, J= 7.2 Hz, 2H), LC-MS: [M+H] 315.1.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2015/196071, 2015, A1 Location in patent: Paragraph 0311

58
[ 2887-98-1 ]
[ 14441-90-8 ]

Yield	Reaction Conditions	Operation in experiment
67%	With 1-hydroxy-3H-benz[d][1,2]iodoxole-1,3-dione In chloroform for 4h; Reflux; Green chemistry;	General Procedure for the Synthesis of Isoxazole-3-carboxylic Acid (6) Ketoxime 5a (0.161 g, 1 mmol), IBX (0.840 g, 3 mmol), and chloroform (5 mL) were placed in a 100 mL round-bottom flask. The reaction was refluxed for 4 h. The reaction was continuously monitored using TLC. The product 6a was purified using column chromatography with petroleum ether and ethyl acetate (9:1) as an eluent. 5-Phenylisoxazole-3-carboxylic acid (6a) [34]: White solid, 67%, mp 162-163 °C; IR (KBr), ν (cm-1): 1450, 1550, 1690, 3000, 3200. 1H NMR (400 MHz, CDCl3), d (ppm): 7.202-8.145 (m, 5H), 9.33(s, 1H). 13C NMR (75 MHz, CDCl3), d (ppm): 30.21, 97.88, 134-124, 142.45, 162.56, 171.54.

Reference： [1]Desai, Vidya G.; Naik, Sneha R.; Dhumaskar, Kashinath L. [Synthetic Communications, 2014, vol. 44, # 10, p. 1453 - 1460]

59
[ 14441-90-8 ]
[ 957793-36-1 ]
ethyl trans-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-1-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
80%	With triethylamine; HATU In tetrahydrofuran at 20℃; for 6h;	9.6 Step 6: ethyl iras-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-l- carboxylate: Ε Ν (5.6 mL, 42 mmol) and HATU (4.84 g, 13 mmol) were added to a mixture of ethyl fras-3-aminocyclobutane-l-carboxylate hydrochloride (1.89 g, 10 mmol) and 5- phenylisoxazole-3-carboxylic acid (2 g, 10 mmol) in THF (200 mL) at room temperature and the reaction mixture was stirred for 6 h at room temperature. Volatiles were removed under reduced pressure to get the crude compound. The reaction mixture was diluted with water (100 mL) and extracted using ethyl acetate (2 x 75 mL). Combined organic layer was washed with brine (50 mL), dried over anhydrous Na2SC>4 and concentrated under reduced pressure. The mixture was purified by flash column chromatography using 30% EtOAc in hexane as eluent to give the product (2.65 g, 80 %) as white solid. Ti NMR (400 MHz, CDC13) : δ 7.80-7.77 (m, 2H), 7.49-7.46 (m, 3H), 7.00 (d, J = 7.2 Hz, 1H), 6.94 (s, 1H), 4.79-4.73 (m, 1H), 4.17 (q, J = 7.1 Hz, 2H), 3.10-3.09 (m, 1H), 2.78-2.72 (m, 2H), 2.40-2.32 (m, 2H), 1.30-1.26 (t, J= 7.2 Hz, 3H). LC-MS: [M+H] + 315.2
80%	With triethylamine; HATU In tetrahydrofuran at 20℃; for 6h;	9.6 ethyl trans-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-1-carboxylate Ε Ν (5.6 mL, 42 mmol) and HATU (4.84 g, 13 mmol) were added to a mixture of ethyl fras-3-aminocyclobutane-l-carboxylate hydrochloride (1.89 g, 10 mmol) and 5- phenylisoxazole-3-carboxylic acid (2 g, 10 mmol) in THF (200 mL) at room temperature and the reaction mixture was stirred for 6 h at room temperature. Volatiles were removed under reduced pressure to get the crude compound. The reaction mixture was diluted with water (100 mL) and extracted using ethyl acetate (2 x 75 mL). Combined organic layer was washed with brine (50 mL), dried over anhydrous Na2S04 and concentrated under reduced pressure. The mixture was purified by flash column chromatography using 30% EtOAc in hexane as eluent to give the product (2.65 g, 80 %) as white solid. Ti NMR (400 MHz, CDC13) : δ 7.80-7.77 (m, 2H), 7.49-7.46 (m, 3H), 7.00 (d, J = 7.2 Hz, 1H), 6.94 (s, 1H), 4.79-4.73 (m, 1H), 4.17 (q, J = 7.1 Hz, 2H), 3.10-3.09 (m, 1H), 2.78-2.72 (m, 2H), 2.40-2.32 (m, 2H), 1.30-1.26 (t, J= 7.2 Hz, 3H). LC-MS: [M+H] + 315.2

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0234
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0193

60
[ 14441-90-8 ]
(5-(cis/trans-3-aminocyclobutyl)-1,3,4-thiadiazol-2-yl)methanol [ No CAS ]
N-cis-(3-(5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]
N-trans-(3-(5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 0.05 g 2: 0.01 g	With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; triethylamine In tetrahydrofuran at 20℃; for 12h;	16.7 Step 7: ^-iras-3-(5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl)cyclobutyl)-5- phenylisoxazole-3-carboxamide and A^-cis-3-(5-(hydroxymethyl)-l,3,4-thiadiazol-2- yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: EDC HCl (0 287 g, 1 mmol), HOBt (0.168 g, 11 mmol) were added to a solution of 5-phenylisoxazole-3-carboxylic acid (0.189 g, 1 mmol) in THF (5 mL), followed by addition of : (5-(3-aminocyclobutyl)-l,3,4-thiadiazol-2- yl)methanol (0.3 g, crude) and the mixture was stirred for 10 min. Triethyl amine (0.42 mL, 3 mmol) was added to the mixture and stir at room temperature for 12h. The reaction mixture was diluted with cold water (20 mL) and extracted with DCM (2 x 10 mL). Combined organic layer was washed with brine and dried over Na2SC>4 and concentrated under reduced pressure to get the crude compound. The crude compound was purified by prep HPLC to afford: A^-cis-3-(5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3- carboxamide (0.05 g): [0278] Yield: 11 % over two steps [0279] Appearance: off white solid. [0280] Analytical data: XH NMR (400 MHz, CDC13): δ 7.83 - 7.79 (m, 2H), 7.53 - 7.49 (m, 3H), 7.22 - 7.20 (br, 1H), 6.97 (s, 1H), 5.08 (s, 2H), 4.75 - 4.68 (m, 1H), 3.77 - 3.68 (m, 1H), 3.10 - 3.03 (m, 2H), 2.62 (br, 1H), 2.57 - 2.51 (m, 2H). [0281] LC-MS: [M+H]+ 356.8 [0282] HPLC purity: 99.19% at 220 nm and 99.11% at 254 nm. A^-iras-3-(5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3- carboxamide (0.01 g): [0283] Yield: 2 % over two steps [0284] Appearance: off white solid. [0285] Analytical data: XH NMR (400 MHz, CDC13): δ 7.80 - 7.78 (m, 2H), 7.50 - 7.47 (m, 3H), 7.11 (d, J= 6.8 Hz, 1H), 6.95 (s, 1H), 5.07 (d, J= 5.4 Hz, 2H), 4.88 - 4.83 (m, 1H), 4.02 - 3.98 (m, 1H), 2.95 - 2.88 (m, 2H), 2.76 - 2.69 (m, 2H), 2.51 - 2.48 (m, 1H). [0286] LC-MS: [M+H]+ 357.1 [0287] HPLC purity: 98.58 % at 220 nm and 98.43 % at 254 nm.
1: 0.01 g 2: 0.05 g	With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; triethylamine In tetrahydrofuran at 20℃; for 12h;	16.7 N-(trans-3-(5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide and N-(cis-3-(5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide EDC.HC1 (0.287 g, 1 mmol), HOBt (0.168 g, 1 1 mmol) were added to a solution of 5-phenylisoxazole-3-carboxylic acid (0.189 g, 1 mmol) in THF (5 mL), followed by addition of : (5-(3-aminocyclobutyl)-l ,3,4-thiadiazol-2- yl)methanol (0.3 g, crude) and the mixture was stirred for 10 min. Triethyl amine (0.42 mL, 3 mmol) was added to the mixture and stir at room temperature for 12h. The reaction mixture was diluted with cold water (20 mL) and extracted with DCM (2 x 10 mL). Combined organic layer was washed with brine and dried over Na2SC>4 and concentrated under reduced pressure to get the crude compound. The crude compound was purified by prep HPLC to afford: A^-cis-3-(5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3- carboxamide (0.05 g): [0237] Yield: 1 1 % over two steps [0238] Appearance: off white solid. [0239] Analytical data: XH NMR (400 MHz, CDC13): δ 7.83 - 7.79 (m, 2H), 7.53 - 7.49 (m, 3H), 7.22 - 7.20 (br, 1H), 6.97 (s, 1H), 5.08 (s, 2H), 4.75 - 4.68 (m, 1H), 3.77 - 3.68 (m, 1H), 3.10 - 3.03 (m, 2H), 2.62 (br, 1H), 2.57 - 2.51 (m, 2H). [0240] LC-MS: [M+H]+ 356.8 [0241] HPLC purity: 99.19% at 220 nm and 99.1 1% at 254 nm. A^-iras-3-(5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3- carboxamide (0.01 g): [0242] Yield: 2 % over two steps [0243] Appearance: off white solid. [0244] Analytical data: lH NMR (400 MHz, CDC13): δ 7.80 - 7.78 (m, 2H), 7.50 - 7.47 (m, 3H), 7.1 1 (d, J = 6.8 Hz, 1H), 6.95 (s, 1H), 5.07 (d, J= 5.4 Hz, 2H), 4.88 - 4.83 (m, 1H), 4.02 - 3.98 (m, 1H), 2.95 - 2.88 (m, 2H), 2.76 - 2.69 (m, 2H), 2.51 - 2.48 (m, 1H). [0245] LC-MS: [M+H]+ 357.1 [0246] HPLC purity: 98.58 % at 220 nm and 98.43 % at 254 nm

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0277-0287
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0236-0245

61
[ 14441-90-8 ]
C₁₄H₂₇N₃OSSi [ No CAS ]
N-((1S,3S)-3-(5-((R)-1-hydroxyethyl)-1,3,4-thiadiazol-2-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
0.11 g	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid; C₁₄H₂₇N₃OSSi With triethylamine; HATU In tetrahydrofuran at 20℃; for 12h; Stage #2: With tetrabutyl ammonium fluoride In tetrahydrofuran for 1h;	19.4; 19.5 Steps 4 and 5: N-((lS,3s)-3-(5-((R)-l-hydroxyethyl)-l,3,4-thiadiazol-2- yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: HATU (0.837 g, 2.2 mmol) was added to a solution of 5-phenylisoxazole-3-carboxylic acid (0.277 g, 1.4 mmol) in THF (5 mL) followed by addition of (R)-l-(5-(3-aminocyclobutyl)-l,3,4-thiadiazol-2-yl)ethan-l-ol (0.46 g, crude) and the resulting reaction mixture was stirred for 10 min. Triethylamine (0.61 mL, 4.4 mmol) was added to the reaction mixture and stirring continued at room temperature for 12h. Cold water (20 mL) was added to the mixture and then extracted with DCM (2 x 10 mL). Combined organic layer was washed with brine, dried over Na2SC>4 and evaporated to dryness under vacuum. The crude compound was dissolved in THF (5 mL) and TBAF solution (1.2 mL, 1.2 mmol) was added and the reaction mixture was stirred for lh. After completion, the reaction mixture was quenched with cold water (20 mL) and extracted with DCM (2 x 5 mL). Combined organic layer was washed with brine and dried over Na2SC>4 and evaporated to dryness under reduced pressure to get the crude compound which was purified by prep HPLC to afford 5-Phenyl-isoxazole-3-carboxylic acid {3-[5-((R)-l-hydroxy-ethyl)-[l,3,4]thiadiazol- 2-yl]-cyclobutyl}-amide (0.110 g, 15 % over two steps) as an off white solid. [0300] Analytical data: XH NMR (400 MHz, CDC13): δ 7.78 - 7.76 (m, 2H), 7.50 - 7.46 (m, 3H), 7. 20 (d, J= 8 Hz, 1H), 6.94 (s, 1H), 5.28 (q, J= 6.5 Hz, 1H), 4.71 - 4.65 (m, 1H), 3.72 - 3.63 (m, 1H), 3.05 - 2.99 (m, 2H), 2.85 (br, 1H), 2.53 - 2.46 (m, 2H), 1.68 (d, J= 6.5Hz, 3H). [0301] LC-MS: [M+H]+ 370.9 [0302] HPLC purity: 98.21% at 220 nm and 98.95% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0299-0302

62
[ 14441-90-8 ]
[ 957793-35-0 ]
ethyl cis-3-(5-phenylisoxazole-3-carboxamido)cyclobutane-1-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
5.02 g	With triethylamine; HATU; In tetrahydrofuran; at 20℃; for 6h;	Et3N (5.3 mL, 0.04 mol) followed by HATU (9.16 g, 0.024 mol) were added to a solution of ethyl czs-3-aminocyclobutane-l-carboxylate hydrochloride (3.63 g, 0.020 mol) and 5-phenylisoxazole-3-carboxylic acid (4.20 g, 0.022 mol) in THF (150 mL) and the reaction mixture was stirred for 6 h at room temperature. Volatiles were removed under reduced pressure and the crude reaction mixture was diluted with water (100 mL). The aq. phase was extracted with ethyl acetate (200 mL). The organic layer was dried over anhydrous Na2SC>4 and concentrated under reduced pressure to obtain the crude product. The crude compound was purified by silica gel column chromatography using 50% EtOAc in hexane as eluent to afford the product (5.02 g, 79 %) as off white solid. XH NMR (400 MHz, CDC13): delta 7.79 -7.77 (m, 2H), 7.50 - 7.46 (m, 3H), 7.08 (d, J = 8.0 Hz, 1H), 6.93 (s, 1H), 4.61 - 4.12 (m, 1H), 4.15 (q, J = 7.1 Hz, 2H), 2.89 - 2.83 (m, 1H), 2.74 - 2.68 (m, 2H), 2.32 - 2.24 (m, 2H), 1.26 (t, J = 7.2 Hz, 2H), LC-MS: [M+H]+ 315.1.
5.02 g	With triethylamine; HATU; In tetrahydrofuran; at 20℃; for 6h;	Et3N (5.3 mL, 0.04 mol) followed by HATU (9.16 g, 0.024 mol) were added to a solution of ethyl czs-3-aminocyclobutane-l-carboxylate hydrochloride (3.63 g, 0.020 mol) and 5-phenylisoxazole-3-carboxylic acid (4.20 g, 0.022 mol) in THF (150 mL) and the reaction mixture was stirred for 6 h at room temperature. Volatiles were removed under reduced pressure and the crude reaction mixture was diluted with water (100 mL). The aq. phase was extracted with ethyl acetate (200 mL). The organic layer was dried over anhydrous Na2SC>4 and concentrated under reduced pressure to obtain the crude product. The crude compound was purified by silica gel column chromatography using 50% EtOAc in hexane as eluent to afford the product (5.02 g, 79 %) as off white solid. l NMR (400 MHz, CDC13): delta 7.79 -7.77 (m, 2H), 7.50 - 7.46 (m, 3H), 7.08 (d, J = 8.0 Hz, 1H), 6.93 (s, 1H), 4.61 - 4.12 (m, 1H), 4.15 (q, J = 7.1 Hz, 2H), 2.89 - 2.83 (m, 1H), 2.74 - 2.68 (m, 2H), 2.32 - 2.24 (m, 2H), 1.26 (t, J = 7.2 Hz, 2H), LC-MS: [M+H]+ 315.1.

Reference： [1]Patent: WO2016/115090,2016,A1 .Location in patent: Paragraph 0305
[2]Patent: WO2017/40606,2017,A1 .Location in patent: Paragraph 0264

63
[ 14441-90-8 ]
1-[1-[trans-3-aminocyclobutyl]-1H-pyrazol-4-yl]ethan-1-ol [ No CAS ]
5-phenyl-N-[trans-3-[4-[(1R)-1-hydroxyethyl]-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]
5-phenyl-N-[trans-3-[4-[(1S)-1-hydroxyethyl]-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 9% 2: 9%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid; 1-[1-[trans-3-aminocyclobutyl]-1H-pyrazol-4-yl]ethan-1-ol With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 25℃; for 2h; Stage #2: In hexane; isopropyl alcohol Resolution of racemate;	26.6 Step 6: 5-phenyl-N-[trans-3-[4-[(lS and lR)-l-hydroxyethyl]-lH-pyrazol-l- yl]cyclobutyl]-l,2-oxazole-3-carboxamide: Into a 50-mL round-bottom flask, was placed a solution of l-[l-[trans-3-aminocyclobut l]-lH-pyrazol-4-yl]ethan-l-ol (226 mg, 1.25 mmol, 1.00 eq.) in DMF (5 mL). To the solution were added 5-phenyl-l,2-oxazole-3-carboxylic acid (282 mg, 1.49 mmol, 1.00 eq.), HATU (700 mg, 1.84 mmol, 1.50 eq.) and DIEA (560 mg, 4.33 mmol, 3.00 eq.). The resulting solution was stirred for 2 hours at 25 °C. The resulting solution was diluted with 100 mL of water. The resulting solution was extracted with ethyl acetate (3x50 mL) and the organic layers combined. The resulting mixture was washed with brine (2x100 mL), dried and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 : 1). The pure isomers were separated by Chiral- Prep-HPLC with the following conditions (Prep-HPLC-004): Column, Phenomenex Lux 5u Cellulose-4 AXIA Packed, 250*21.2mm,5um; mobile phase, Hex and IPA (hold 50.0% IPA in 15 min); Detector, UV 254/220nm. This resulted in 39.6 mg (9%) of 5-phenyl-N-[trans-3-[4- [(lR)-l-hydroxyethyl]-lH-pyrazol-l-yl]cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid and 39.4 mg (9%) of 5-phenyl-N-[trans-3-[4-[(l S)-l-hydroxyethyl]-lH-pyrazol-l- yl]cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid: [0340] Isomer 1: [0341] Analytical data: XH NMR (300 MHz, OMSO-d6): δ 9.31-9.28 (d, J= 7.2 Hz, 2H), 7.96-7.92 (m. 2H), 7.68 (s, 1H), 7.59-7.55 (m, 3H), 7.41 (s, 1H), 7.38 (s, 1H), 5.00-4.89 (m, 1H), 4.88-4.86 (d, J= 4.8Hz, 1H), 4.71-4.64 (m, 2H), 2.76-2.61 (m, 4H), 1.34-1.32 (d, J = 6.3 Hz, 3H). [0342] LC-MS: (M+H)+ = 353 [0343] HPLC purity: 99.24 at 254 nm [0344] Isomer 2: [0345] Analytical data: lH NMR (300 MHz, OMSO-d6): δ 9.31-9.28 (d, J = 7.5 Hz, 2H), 7.96-7.93 (m, 2H), 7.68 (s, 1H), 7.57-7.55 (m, 3H), 7.41 (s, 1H), 7.38 (s, 1H), 4.97-4.89 (m, 1H), 4.88-4.86 (d, J= 4.8 Hz, 1H), 4.70-4.64 (m, 2H), 2.72-2.61 (m, 4H), 1.34-1.32 (d, J = 6.6 Hz, 3H). [0346] LC-MS: (M+H)+ = 353 [0347] HPLC purity: 99.74 at 254 nm.
1: 39.6 mg 2: 39.4 mg	With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 25℃; for 2h;	26.6 5-phenyl-N-[trans-3-[4-[(lS and lR)-l-hydroxyethyl]-lH-pyrazol-l- yl]cyclobutyl]-l,2-oxazole-3-carboxamide Into a 50-mL round-bottom flask, was placed a solution of l-[l-[trans-3-aminocyclobu†yl]-lH-pyrazol-4-yl]ethan-l-ol (226 mg, 1.25 mmol, 1.00 eq.) in DMF (5 mL). To the solution were added 5-phenyl-l,2-oxazole-3-carboxylic acid (282 mg, 1.49 mmol, 1.00 eq.), HATU (700 mg, 1.84 mmol, 1.50 eq.) and DIEA (560 mg, 4.33 mmol, 3.00 eq.). The resulting solution was stirred for 2 hours at 25 °C. The resulting solution was diluted with 100 mL of water. The resulting solution was extracted with ethyl acetate (3x50 mL) and the organic layers combined. The resulting mixture was washed with brine (2x100 mL), dried and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 : 1). The pure isomers were separated by Chiral- Prep-HPLC with the following conditions (Prep-HPLC-004): Column, Phenomenex Lux 5u Cellulose-4 AXIA Packed, 250*21.2mm,5um; mobile phase, Hex and IPA (hold 50.0% IPA in 15 min); Detector, UV 254/220nm. This resulted in 39.6 mg (9%) of 5-phenyl-N-[trans-3-[4- [(lR)-l-hydroxyethyl]-lH-pyrazol-l-yl]cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid and 39.4 mg (9%) of 5-phenyl-N-[trans-3-[4-[(l S)-l-hydroxyethyl]-lH-pyrazol-l- yl]cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid: [0299] Isomer 1: [0300] Analytical data: lH NMR (300 MHz, OMSO-d6): δ 9.31-9.28 (d, J= 7.2 Hz, 2H), 7.96-7.92 (m. 2H), 7.68 (s, 1H), 7.59-7.55 (m, 3H), 7.41 (s, 1H), 7.38 (s, 1H), 5.00-4.89 (m, 1H), 4.88-4.86 (d, J= 4.8Hz, 1H), 4.71-4.64 (m, 2H), 2.76-2.61 (m, 4H), 1.34-1.32 (d, J = 6.3 Hz, 3H). [0301] LC-MS: (M+H)+ = 353 [0302] HPLC purity: 99.24 at 254 nm [0303] Isomer 2: [0304] Analytical data: XH NMR (300 MHz, DMSO-c): δ 9.31-9.28 (d, J = 7.5 Hz, 2H), 7.96-7.93 (m, 2H), 7.68 (s, 1H), 7.57-7.55 (m, 3H), 7.41 (s, 1H), 7.38 (s, 1H), 4.97-4.89 (m, 1H), 4.88-4.86 (d, J= 4.8 Hz, 1H), 4.70-4.64 (m, 2H), 2.72-2.61 (m, 4H), 1.34-1.32 (d, J = 6.6 Hz, 3H). [0305] LC-MS: (M+H)+ = 353 [0306] HPLC purity: 99.74 at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0339
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0298-0305

64
[ 14441-90-8 ]
[1-[cis-3-aminocyclobutyl]-1H-pyrazol-4-yl]methanol hydrochloride [ No CAS ]
5-phenyl-N-[cis-3-[4-(hydroxymethyl)-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
63 mg	With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 20℃; for 1h;	28.3 Step 3: 5-phenyl-N-[cis-3-[4-(hydroxymethyl)-lH-pyrazol-l-yl]cyclobutyl]-l,2- oxazole-3-carboxamide: into a 50-mL round-bottom flask, was placed a solution of [l-[czs-3- aminocyclobutyl]-lH-pyrazol-4-yl]methanol hydrochloride (408 mg, 2.00 mmol, 1.20 eq.) in DMF (10 mL). To the solution were added DIEA (645 mg, 4.99 mmol, 3.00 eq.), 5-phenyl- l,2-oxazole-3-carboxylic acid (315 mg, 1.67 mmol, 1.00 eq.) and HATU (760 mg, 2.00 mmol, 1.20 eq.). The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of 20 mL of water. The resulting solution was extracted with ethyl acetate (3x20 mL) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a Prep-TLC with ethyl acetate/petroleum ether (2: 1). This resulted in 63 mg (11%) of 5-phenyl-N-[cw-3-[4- (hydroxy methyl)- 1 H-py razol- 1 -y 1] cy clobuty 1] - 1 ,2-oxazole-3 -carboxamide. [0366] Appearance: white solid [0367] Analytical data: 1HNMR (400MHz, DMSO-i): δ 9.24, 9.22 (d, J = 8.0Hz, 1H), 7.95-7.92 (m, 2H), 7.80 (s, 1H), 7.59-7.53 (m, 3H), 7.38, 7.36 (d, J = 8.0Hz, 2H), 4.83-4.80 (t, J = 6.0Hz, 1H), 4.60-4.56 (m, 1H), 4.36-4.28 (m, 3H), 2.82-2.75 (m, 2H), 2.65-2.59 (m, 2H). [0368] LC-MS: (M+H)+ = 339 [0369] HPLC purity: 99.94% at 254 nm.
63 mg	With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 20℃; for 1h;	28.3 5-phenyl-N-[cis-3-[4-(hydroxymethyl)-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide into a 50-mL round-bottom flask, was placed a solution of [l-[czs-3- aminocyclobutyl]-lH-pyrazol-4-yl]methanol hydrochloride (408 mg, 2.00 mmol, 1.20 eq.) in DMF (10 mL). To the solution were added DIEA (645 mg, 4.99 mmol, 3.00 eq.), 5-phenyl- l,2-oxazole-3-carboxylic acid (315 mg, 1.67 mmol, 1.00 eq.) and HATU (760 mg, 2.00 mmol, 1.20 eq.). The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of 20 mL of water. The resulting solution was extracted with ethyl acetate (3x20 mL) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a Prep-TLC with ethyl acetate/petroleum ether (2: 1). This resulted in 63 mg (11%) of 5-phenyl-N-[cw-3-[4- (hydroxy methyl)- 1 H-py razol- 1 -y 1] cy clobuty 1] - 1 ,2-oxazole-3 -carboxamide. [0325] Appearance: white solid [0326] Analytical data: 1HNMR (400MHz, DMSO-i): δ 9.24, 9.22 (d, J = 8.0Hz, 1H), 7.95-7.92 (m, 2H), 7.80 (s, 1H), 7.59-7.53 (m, 3H), 7.38, 7.36 (d, J = 8.0Hz, 2H), 4.83-4.80 (t, J = 6.0Hz, 1H), 4.60-4.56 (m, 1H), 4.36-4.28 (m, 3H), 2.82-2.75 (m, 2H), 2.65-2.59 (m, 2H). [0327] LC-MS: (M+H)+ = 339 [0328] HPLC purity: 99.94% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0365-0369
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0324-0328

65
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[1-[cis-3-aminocyclobutyl]-1H-pyrazol-3-yl]methanol hydrochloride [ No CAS ]
[1-[cis-3-(5-phenyl-1,2-oxazole-3-carboxamido)cyclobutyl]-1H-pyrazol-3-yl]methyl 5-phenyl-1,2-oxazole-3-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
40%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 2h;	30.4 Step 4: [l-[cis-3-(5-phenyl-l,2-oxazole-3-amido)cyclobutyl]-lH-pyrazol-3- yljmethyl 5-phenyl-l,2-oxazole-3-carboxylate: into a 100-mL round-bottom flask, was placed a solution of [l-fcw-S-aminocyclobutylJ-lH-pyrazol-S-yllmethanol hydrochloride (400 mg, 1.96 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (745 mg, 3.94 mmol, 2.00 eq.), HCTU (983 mg, 2.36 mmol, 1.20 eq.) and DIEA (762 mg, 5.90 mmol, 3.00 eq.) in DMF (20 mL). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with ethyl acetate and the organic layers combined. The resulting mixture was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The crude residue was washed with 5 mL of methanol. This resulted in 400 mg (40%) of [l-[czs-3-(5-phenyl-l,2-oxazole-3- amido)cyclobutyl]-lH-pyrazol-3-yl]methyl 5-phenyl-l,2-oxazole-3-carboxylate as a white solid. LC-MS: (M+H)+ = 510.
40%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 2h;	30.4 [1-[cis-3-(5-phenyl-1,2-oxazole-3-amido)cyclobutyl]-1H-pyrazol-3-yl]methyl 5-phenyl-1,2-oxazole-3-carboxylate into a 100-mL round-bottom flask, was placed a solution of [l-fcw-S-aminocyclobutylJ-lH-pyrazol-S-yllmethanol hydrochloride (400 mg, 1.96 mmol, 1.00 eq.), 5-phenyl-l,2-oxazole-3-carboxylic acid (745 mg, 3.94 mmol, 2.00 eq.), HCTU (983 mg, 2.36 mmol, 1.20 eq.) and DIEA (762 mg, 5.90 mmol, 3.00 eq.) in DMF (20 mL). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with ethyl acetate and the organic layers combined. The resulting mixture was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The crude residue was washed with 5 mL of methanol. This resulted in 400 mg (40%) of [l-[czs-3-(5-phenyl-l,2-oxazole-3- amido)cyclobutyl]-lH-pyrazol-3-yl]methyl 5-phenyl-l,2-oxazole-3-carboxylate as a white solid. LC-MS: (M+H)+ = 510.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0376
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0335

66
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1-[1-[trans-3-aminocyclobutyl]-1H-pyrazol-3-yl]ethan-1-ol hydrochloride [ No CAS ]
5-phenyl-N-[trans-3-[5-(1-hydroxyethyl)-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
57%	With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 20℃; for 2h;	27.3 Step 3: 5-phenyl-At/iras-3-[3-(l-hydroxyethyl)-lH-pyrazol-l-yl]cyclobutyl]- l,2-oxazole-3-carboxamide: into a 50-mL round-bottom flask, was placed a solution of 5- - I l l - phenyl-l,2-oxazole-3-carboxylic acid (177.7 mg, 0.94 mmol, 1.00 eq.), l-[l-[trans-3- aminocyclobutyl]-lH-pyrazol-3-yl]ethan-l-ol hydrochloride (246 mg, 1.13 mmol, 1.20 eq.), HATU (428.8 mg, 1.13 mmol, 1.20 eq.) and DIEA (363.9 mg, 2.82 mmol, 3.00 eq.) in DMF(10 mL). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of 20 mL of water. The resulting solution was extracted with ethyl acetate (3x20 mL) and the organic combined layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a Prep-TLC with ethyl acetate/petroleum ether (2: 1). This resulted in 189 mg (57%) of 5-pheny-N-[trans-3-[3-(- hydroxyethyl)-lH-pyrazol-l-yl]cyclobut l]-l,2-oxazole-3-carboxamide as an off-white solid.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0350

67
[ 14441-90-8 ]
[[1-trans-3-aminocyclobutyl]-1H-pyrazol-3-yl]methanol [ No CAS ]
5-phenyl-N-[trans-3-[3-(hydroxymethyl)-1H-pyrazol-1-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
18%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃;	31 Example 31: N-iras-3-(3-(hydroxymethyl)-lH-pyrazol-l-yl)cyclobutyl)-5- phenylis oxazole-3-carb oxamide Into a 100-mL round-bottom flask, was placed a solution of [ -trans-3- aminocyclobutyl]-lH-pyrazol-3-yl]methanol (120 mg, 0.72 mmol, 1.00 eq., prepared using similar procedure as shown in example 29) in dichloromethane (5 mL). To the solution were added 5-phenyl-l ,2-oxazole-3-carboxylic acid (163 mg, 0.86 mmol, 1.20 eq.) and HCTU (360 mg, 0.87 mmol, 1.20 eq.). This was followed by the addition of DIEA (278 mg, 2.15 mmol, 3.00 eq.) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with dichloromethane (3x50 mL). The organic layers were combined, dried and concentrated under vacuum. The crude product was purified by Prep-HPLC with the following conditions (Waters): Column, Bridget Prep C18 5um OBDTM 19* 100mm; mobile phase, water with 0.05% NH4HCO3 and CH3CN (40.0% CH3CN up to 80.0% in 10 min, up to 95.0% in 1.5min, down to 40.0% in 1.5min); Detector, 254nm. This resulted in 44.7 mg (18%) of 5- phenyl-N-frara-S-fS-^ydroxymethy^-lH-pyrazol-l-ylJcyclobutylJ-l^-oxazole-S-carboxamide as a white solid. [0382] LC-MS: (M+H)+ = 339 [0383] Analytical data: lH NMR (400MHz, DMSO-i): δ 9.32-9.30 (d, J= 6.8 Hz, 1H), 7.95-7.94 (d, J = 6.0 Hz, 2H), 7.74 (s, 1H), 7.57-7.55 (m, 3H), 7.38 (s, 1H), 6.20 (s, 1H), 5.02- 4.99 (t, J= 5.6Hz, 1H), 4.96-4.95 (m, 1H), 4.71-4.65 (m, 1H), 4.44-4.42 (d, J= 6.0 Hz, 2H), 2.75-2.63 (m, 4H). [0384] HPLC purity: 98.8% at 254 nm.
18%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 1h;	31 N-(trans-3-(3-(hydroxymethyl)-1H-pyrazol-1-yl)cyclobutyl)-5-phenylisoxazole-3-carboxamide Into a 100-mL round-bottom flask, was placed a solution of [l -trans-3- aminocyclobutyl]-lH-pyrazol-3-yl]methanol (120 mg, 0.72 mmol, 1.00 eq., prepared using similar procedure as shown in example 29) in dichloromethane (5 mL). To the solution were added 5-phenyl-l ,2-oxazole-3-carboxylic acid (163 mg, 0.86 mmol, 1.20 eq.) and HCTU (360 mg, 0.87 mmol, 1.20 eq.). This was followed by the addition of DIEA (278 mg, 2.15 mmol, 3.00 eq.) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with dichloromethane (3x50 mL). The organic layers were combined, dried and concentrated under vacuum. The crude product was purified by Prep-HPLC with the following conditions (Waters): Column, Bridget Prep C18 5um OBDTM 19* 100mm; mobile phase, water with 0.05% NH4HC03 and CH3CN (40.0% CH3CN up to 80.0% in 10 min, up to 95.0% in 1.5min, down to 40.0% in 1.5min); Detector, 254nm. This resulted in 44.7 mg (18%) of 5- phenyl-N-frara-S-fS-^ydroxymethy^-lH-pyrazol-l-ylJcyclobutylJ-l^-oxazole-S-carboxamide as a white solid. [0341] LC-MS: (M+H)+ = 339 [0342] Analytical data: XH NMR (400MHz, DMSO-i): δ 9.32-9.30 (d, J= 6.8 Hz, 1H), 7.95-7.94 (d, J = 6.0 Hz, 2H), 7.74 (s, 1H), 7.57-7.55 (m, 3H), 7.38 (s, 1H), 6.20 (s, 1H), 5.02- 4.99 (t, J= 5.6Hz, 1H), 4.96-4.95 (m, 1H), 4.71-4.65 (m, 1H), 4.44-4.42 (d, J= 6.0 Hz, 2H), 2.75-2.63 (m, 4H). [0343] HPLC purity: 98.8% at 254 nm

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0381-0384
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0340-0343

68
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[5-(3-aminocyclobutyl)-1,2,4-oxadiazol-3-yl]methanol [ No CAS ]
5-phenyl-N-[cis-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]
5-phenyl-N-[trans-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 37% 2: 3%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃;	32.7; 32.8 Step 7: iV-[3-[3-(hydroxymethyl)-l,2,4-oxadiazol-5-yl]cyclobutyl]-5-phenyl-l,2- oxazole-3-carboxamide: into a 250-mL round-bottom flask, was placed a solution of 5-phenyl- l,2-oxazole-3-carboxylic acid (1 g, 5.28 mmol, 1.10 eq.) in dichloromethane (100 mL). To the mixture were added [5-(3-aminocyclobutyl)-l,2,4-oxadiazol-3-yl]methanol (800 mg, 4.52 mmol, 1.00 eq.) and HATU (2.16 g, 8.96 mmol, 1.20 eq.). This was followed by the addition of DIEA (2 g, 14.5 mmol, 3.00 eq.) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with dichloromethane (3x100 mL) and the organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with dichloromethane/methanol (50: 1). This resulted in 1 g (62%) of N-[3-[3-(hydroxymethyl)-l,2,4-oxadiazol-5-yl]cyclobut l]-5-phenyl-l,2-oxazole-3- carboxamide as a white solid. LC-MS: [M+H]+ = 341. [0393] Step 8: Separation by SFC: the isomers (lg) were separated by Chiral-Prep-HPLC with the following conditions (Prep-HPLC-032): Column, Repaired IA, 21.2* 150mm, 5um; mobile phase, Hex and ethanol (hold 50.0% ethanol in 15 min); Detector, UV 254/220nm. This resulted in 555 mg (37%) of 5-p my-N-[trans-3-[3-( ydroxymQt y)-,2,4-oxadiazo-5- yl]cyclobutyl]-l,2-oxazole-3-carboxamide and 26.5 mg (3%) of 5-phenyl-N-[cw-3-[3- (hydroxymethyl)-l,2,4-oxadiazol-5-yl]cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid. [0394] 5-Phenyl-^-[iras-3-[3-(hydroxymethyl)-l,2,4-oxadiazol-5-yl]cyclobutyl]-l,2- oxazole-3-carboxamide [0395] Analytical data: XH NMR (300MHz, DMSO-i): δ 9.34-9.31 (d, J= 7.8 Hz, 1H), 7.96-7.93 (m, 2H), 7.78-7.75 (m, 3H), 7.60-7.55 (m, 3H), 7.38 (s, 1H), 5.72-5.68 (t, J = 6.3 Hz, 1H), 4.81-4.70 (m, 1H), 4.57-4.55 (d, J = 6.3 Hz, 2H), 3.81-3.75 (m, 1H), 2.78-2.71 (m, 2H), 2.68-2.58 (m, 2H). [0396] HPLC purity: 99.27% at 254 nm. [0397] 5-Phenyl-^-[cis-3-[3-(hydroxymethyl)-l,2,4-oxadiazol-5-yl]cyclobutyl]-l,2- oxazole-3-carboxamide [0398] Analytical data: ^-NMR (400MHz, CD3OD-c): δ 7.91-7.88 (m, 2H), 7.57-7.52 (m, 3H), 7.10 (s, 1H), 4.72-4.60 (m, 3H), 3.73-3.58 (m, 1H), 2.91-2.87 (m, 2H), 2.65-2.57 (m, 2H). [0399] HPLC purity: 98.2% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0392-0399

69
[ 14441-90-8 ]
2-(3-aminocyclobutyl)ethan-1-ol hydrochloride [ No CAS ]
5-phenyl-N-[trans-3-(2-hydroxyethyl)cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]
5-phenyl-N-[cis-3-(2-hydroxyethyl)cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 60% 2: 70%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	36.2 Step 2: N-[3-(2-hydroxyethyl)cyclobutyl]-5-phenyl-l,2-oxazole-3-carboxamide: a solution of 5-phenyl-l,2-oxazole-3-carboxylic acid (850.5 mg, 4.50 mmol, 1.51 eq.) and 2-(3- aminocyclobutyl)ethan-l-ol hydrochloride (452 mg, 2.98 mmol, 1.00 eq.) in dichloromethane (25 mL)was placed in a 100-mL round-bottom flask. HATU (1.368 g, 3.60 mmol, 1.21 eq.) and DIEA (1.161 g, 8.98 mmol, 3.01 eq.) were added to the solution and stirred for 1 hour at room temperature. The resulting solution was diluted with 50 mL of water, extracted with chloromethane (3x30 mL) and the organic layers combined. The resulting mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC with the following conditions (CombiFlash-1): Column, C18 silica gel; mobile phase, MeCN/H20=55:45 increasing to MeCN/H2O=60:40 within 2 min; Detector, UV 254 nm to give 110 mg (13%) of N-[3-(2-hydroxyethyl)cyclobutyl]-5-phenyl- l,2-oxazole-3-carboxamide as a off-white solid. The isomers were separated by Chiral-Prep- HPLC using the following conditions (Prep-HPLC-009): Column, Repaired IA, 21.2* 150mm, 5um; mobile phase, Hexane and ethanol (hold 20.0% ethanol in 20 min); Detector, UV 254/220 nm. This resulted in 23.8 mg (60%) of 5-phenyl-N-[ ram,-3-(2-hydroxyethyl)cyclobutyl]-l,2- oxazole-3-carboxamide as a white solid and 35.7 mg (70%) of 5-phenyl-N-[cw-3-(2- hydroxyethyl)cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid. iV-iras-3-(2-hydroxyethyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: [0439] LC-MS: (M+H)+ = 287 [0440] Analytical data: XH NMR (CDC13, 400MHz): δ 7.83-7.81 (m, 2H), 7.54-7.49 (m, 3H), 7.05-7.02 (m, 1H), 6.97 (s, 1H), 4.72-4.67 (m, 1H), 3.73-3.68 (t, J = 10.0Hz, 2H), 2.49- 2.41 (m, 1H), 2.26-2.21 (m, 4H), 1.87-1.81 (m, 2H). [0441] HPLC purity: 99.4% at 254 nm. iV-cis-3-(2-hydroxyethyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: [0442] LC-MS: (M+H)+ = 287 [0443] Analytical data: XH NMR (CDC13, 400MHz): 7.83-7.80 (m, 2H), 7.54-7.49 (m, 3H), 7.02-6.93 (m, 3H), 4.50-4.44 (m, 1H), 3.67-3.63 (t, J= 8.0Hz, 2H), 2.68-2.62 (m, 2H), 2.22- 2.13 (m, 1H), 1.76-1.66 (m, 4H). [0444] HPLC purity: 99.0% at 254 nm.
1: 23.8 mg 2: 35.7 mg	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	36.2 N-[3-(2-hydroxyethyl)cyclobutyl]-5-phenyl-l,2-oxazole-3-carboxamide a solution of 5-phenyl-l,2-oxazole-3-carboxylic acid (850.5 mg, 4.50 mmol, 1.51 eq.) and 2-(3- aminocyclobutyl)ethan-l-ol hydrochloride (452 mg, 2.98 mmol, 1.00 eq.) in dichloromethane (25 mL)was placed in a 100-mL round-bottom flask. HATU (1.368 g, 3.60 mmol, 1.21 eq.) and DIEA (1.161 g, 8.98 mmol, 3.01 eq.) were added to the solution and stirred for 1 hour at room temperature. The resulting solution was diluted with 50 mL of water, extracted with chloromethane (3x30 mL) and the organic layers combined. The resulting mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC with the following conditions (CombiFlash-1): Column, C18 silica gel; mobile phase, MeCN/H20=55:45 increasing to MeCN/H2O=60:40 within 2 min; Detector, UV 254 nm to give 110 mg (13%) of N-[3-(2-hydroxyethyl)cyclobutyl]-5-phenyl- l,2-oxazole-3-carboxamide as a off-white solid. The isomers were separated by Chiral-Prep- HPLC using the following conditions (Prep-HPLC-009): Column, Repaired IA, 21.2* 150mm, 5um; mobile phase, Hexane and ethanol (hold 20.0% ethanol in 20 min); Detector, UV 254/220 nm. This resulted in 23.8 mg (60%) of 5-phenyl-N-[ ram,-3-(2-hydroxyethyl)cyclobutyl]-l,2- oxazole-3-carboxamide as a white solid and 35.7 mg (70%) of 5-phenyl-N-[cw-3-(2- hydroxyethyl)cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid. iV-iras-3-(2-hydroxyethyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: [0393] LC-MS: (M+H)+ = 287 [0394] Analytical data: XH NMR (CDC13, 400MHz): δ 7.83-7.81 (m, 2H), 7.54-7.49 (m, 3H), 7.05-7.02 (m, 1H), 6.97 (s, 1H), 4.72-4.67 (m, 1H), 3.73-3.68 (t, J = 10.0Hz, 2H), 2.49- 2.41 (m, 1H), 2.26-2.21 (m, 4H), 1.87-1.81 (m, 2H). [0395] HPLC purity: 99.4% at 254 nm. iV-cis-3-(2-hydroxyethyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide: [0396] LC-MS: (M+H)+ = 287 [0397] Analytical data: XH NMR (CDC13, 400MHz): 7.83-7.80 (m, 2H), 7.54-7.49 (m, 3H), 7.02-6.93 (m, 3H), 4.50-4.44 (m, 1H), 3.67-3.63 (t, J= 8.0Hz, 2H), 2.68-2.62 (m, 2H), 2.22- 2.13 (m, 1H), 1.76-1.66 (m, 4H). [0398] HPLC purity: 99.0% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0438-0444
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0392-0398

70
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cis-3-aminocyclobutanecarbonitrile hydrochloride [ No CAS ]
N-(cis-3-cyanocyclobutyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
600 mg	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	37.3 Step 3: iV-(ciV3-cyanocyclobutyl)-5-phenylisoxazole-3-carboxamide: 3- aminocyclobutanecarbonitrile hydrochloride (440 mg, 4.58 mmol, 1.00 eq.), 5-phenyl-l,2- oxazole-3-carboxylic acid (866 mg, 4.58 mmol, 1.00 eq.) and HATU (2090 mg, 5.50 mmol, 1.20 eq.) in dichloromethane (18 mL) were placed in a 100-mL round-bottom flask. To the mixture was added DIEA (1773 mg, 13.72 mmol, 3.00 eq.) and the mixture was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with ethyl acetate and the organic layers combined. The resulting mixture was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 :3) to give 600 mg (49%) of N-(cw-3-cyanocyclobutyl)-5- phenylisoxazole-3-carboxamide as a white solid. LC-MS: (M+H)+ = 268.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0447

71
[ 14441-90-8 ]
3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutan-1-amine [ No CAS ]
5-phenyl-N-[cis-3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]
5-phenyl-N-[trans-3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 13% 2: 59%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	38.1 Stepl: 5-phenyl-^-[cis-3-(3-[l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,2,4- oxadiazol-5-yl)cyclobutyl]-l,2-oxazole-3-carboxamide and 5-phenyl-iV-[iras-3-(3-[l- [(tert-butyldimethylsilyl)oxy]ethyl]-l,2,4-oxadiazol-5-yl)cyclobutyl]-l,2-oxazole-3- carboxamide: to a solution of 3-(3-[l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,2,4-oxadiazol-5- yl)cyclobutan-l -amine (1.5 g, 5.04 mmol, 1.00 eq.) in dichloromethane (100 mL)was added 5- phenyl-l,2-oxazole-3-carboxylic acid (1.13 g, 5.97 mmol, 1.20 eq.), HATU (2.28 g, 6.00 mmol, 1.20 eq.) and DIEA (1.93 g, 14.93 mmol, 3.00 eq.). The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water and extracted with ethyl acetate (3x50 mL) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (l :50)to give 300 mg (13%) of 5-phenyl- N-[cw-3-(3-[l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,2,4-oxadiazol-5-yl)cyclobutyl]-l,2- oxazole-3-carboxamide as a white solid. The solvent was changed to a mixture of with ethyl acetate/petroleum ether (1 :20) to give 1.4 g (59%) of 5-phenyl-N-[fra ,-3-(3-[l-[(tert- butyldimethylsilyl)oxy]ethyl]-l,2,4-oxadiazol-5-yl)cyclobutyl]-l,2-oxazole-3-carboxamide as a white solid. LC-MS: (M+H)+ = 469.
1: 1.4 g 2: 300 mg	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 1h;	38.1 5-phenyl-N-[cis-3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutyl]-1,2-oxazole-3-carboxamide and 5-phenyl-N-[trans-3-(3-[1-[(tert-butyldimethylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)cyclobutyl]-1,2-oxazole-3-carboxamide to a solution of 3-(3-[l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,2,4-oxadiazol-5- yl)cyclobutan-l -amine (1.5 g, 5.04 mmol, 1.00 eq.) in dichloromethane (100 mL)was added 5- phenyl-l,2-oxazole-3-carboxylic acid (1.13 g, 5.97 mmol, 1.20 eq.), HATU (2.28 g, 6.00 mmol, 1.20 eq.) and DIEA (1.93 g, 14.93 mmol, 3.00 eq.). The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water and extracted with ethyl acetate (3x50 mL) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 :50)to give 300 mg (13%) of 5-phenyl- N-[cw-3-(3-[l-[(tert-butyldimethylsilyl)oxy]ethyl]-l,2,4-oxadiazol-5-yl)cyclobu†yl]-l,2- oxazole-3-carboxamide as a white solid. The solvent was changed to a mixture of with ethyl acetate/petroleum ether (1 :20) to give 1.4 g (59%) of 5-phenyl-N-[fra ,-3-(3-[l-[(tert- bu†yldimethylsilyl)oxy]ethyl]-l,2,4-oxadiazol-5-yl)cyclobu†yl]-l,2-oxazole-3-carboxamide as a white solid. LC-MS: (M+H)+ = 469

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0456
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0410

72
[ 14441-90-8 ]
[5-[(3-aminocyclobutyl)methyl]-1,3,4-thiadiazol-2-yl]methanol hydrogen chloride salt [ No CAS ]
5-phenyl-N-[cis-3-[[5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl]methyl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]
5-phenyl-N-[trans-3-[[5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl]methyl]cyclobutyl]-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
1: 37% 2: 37%	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 2h;	5.3 Step 3: N-(iras-3-((5-(hydroxymethyl)-l,3,4-thiadiazol-2- yl)methyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide and N-(cis-3-((5- (hydroxymethyl)-l,3,4-thiadiazol-2-yl)methyl)cyclobutyl)-5-phenylisoxazole-3- carboxamide: a solution of [5-[(3-aminocyclobutyl)methyl]-l,3,4-thiadiazol-2-yl]methanol hydrochloride (500 mg, 2.12 mmol, 1.00 eq., 99%), 5-phenyl-l,2-oxazole-3-carboxylic acid (481 mg, 2.54 mmol, 1.20 eq.), HCTU (1.061 g, 2.55 mmol, 1.20 eq.) and DIEA (1.09 g, 8.43 mmol, 1.20 eq.) in dichloromethane (30 mL) was stirred for 2 hours at room temperature. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep- Flash with acetonitrile and water (0-46% within 40 min). The isomers were separated by Prep- SFC with the following conditions (prep SFC 350-2): Column, Phenomenex Lux 5μ Cellulose- 4, 250*50mm; mobile phase, C02 (50%), MeOH (0.2%DEA) (50%); Detector, UV 220nm. 5-phenyl-iV- [(ci's-3- [ [5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl] methyl] cyclobutyl] - 1,2- oxazole-3-carboxamide: [0198] Yield: 37% [0199] Appearance: off-white solid [0200] Analytical data: XH NMR (400MHz, OMSO-d6, ppm): δ: 9.06 (d, J= 8.0Hz, 1H), 7.94-7.92 (m, 2H), 7.58-7.54 (m, 3H), 7.35 (s, 1H), 6.14-6.11 (m, 1H), 4.80 (d, J = 6.0Hz, 2H), 4.35-4.33 (m, 1H), 3.19-3.17 (m, 2H), 2.43-2.33 (m, 3H), 1.99-1.93 (m, 2H). [0201] LC-MS: 371.1 [M+H]+ 5-phenyl-iV- [(trans-3- [ [5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl] methyl] cyclobutyl] - 1,2- oxazole-3-carboxamide: [0202] Yield: 37% [0203] Appearance: light yellow solid [0204] Analytical data: NMR (400MHz, OMSO-d6, ppm): δ: 9.14 (d, J= 7.2Hz, 1H), 7.94-7.92 (m, 2H), 7.56-7.54 (m, 3H), 7.36 (s, 1H), 6.14-6.11 (m, 1H), 4.80 (d, J= 6.0Hz, 2H), 4.63-4.55 (m, 1H), 3.33-3.28 (m, 2H), 2.51-2.49 (m, 1H), 2.33-2.31 (m, 2H) , 2.14-2.13 (m, 2H).
	With 6-chloro-3-((dimethylamino)(dimethyliminio)methyl)-1H-benzo[d][1,2,3]triazol-3-ium-1-olatehexafluorophosphate(V); N-ethyl-N,N-diisopropylamine In dichloromethane at 20℃; for 2h;	5.3 N-(trans-3-((5-(hydroxymethyl)-l,3,4-thiadiazol-2- yl)methyl)cyclobutyl)-5-phenylisoxazole-3-carboxamide and N-(cis-3-((5- (hydroxymethyl)- 1,3,4- thiadiazol-2-yl)methyl)cyclobutyl)-5-phenylisoxazole-3- carboxamide a solution of [5-[(3-aminocyclobutyl)methyl]-l,3,4-thiadiazol-2-yl]methanol hydrochloride (500 mg, 2.12 mmol, 1.00 eq., 99%), 5-phenyl-l,2-oxazole-3-carboxylic acid (481 mg, 2.54 mmol, 1.20 eq.), HCTU (1.061 g, 2.55 mmol, 1.20 eq.) and DIEA (1.09 g, 8.43 mmol, 1.20 eq.) in dichloromethane (30 mL) was stirred for 2 hours at room temperature. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep- Flash with acetonitrile and water (0-46% within 40 min). The isomers were separated by Prep- SFC with the following conditions (prep SFC 350-2): Column, Phenomenex Lux 5μ Cellulose- 4, 250*50mm; mobile phase, C02 (50%), MeOH (0.2%DEA) (50%); Detector, UV 220nm. 5-phenyl-iV- [(cis-3- [ [5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl] methyl] cyclobutyl] - 1,2- oxazole-3-carboxamide: [0157] Yield: 37% [0158] Appearance: off-white solid [0159] Analytical data: XH NMR (400MHz, OMSO-d6, ppm): δ: 9.06 (d, J= 8.0Hz, 1H), 7.94-7.92 (m, 2H), 7.58-7.54 (m, 3H), 7.35 (s, 1H), 6.14-6.11 (m, 1H), 4.80 (d, J= 6.0Hz, 2H), 4.35-4.33 (m, 1H), 3.19-3.17 (m, 2H), 2.43-2.33 (m, 3H), 1.99-1.93 (m, 2H). [0160] LC-MS: 371.1 [M+H]+ 5-phenyl-iV- [(trans-3- [ [5-(hydroxymethyl)-l,3,4-thiadiazol-2-yl] methyl] cyclobutyl] - 1,2- oxazole-3-carboxamide: [0161] Yield: 37% [0162] Appearance: light yellow solid [0163] Analytical data: NMR (400MHz, OMSO-d6, ppm): δ: 9.14 (d, J= 7.2Hz, 1H), 7.94-7.92 (m, 2H), 7.56-7.54 (m, 3H), 7.36 (s, 1H), 6.14-6.11 (m, 1H), 4.80 (d, J= 6.0Hz, 2H), 4.63-4.55 (m, 1H), 3.33-3.28 (m, 2H), 2.51-2.49 (m, 1H), 2.33-2.31 (m, 2H) , 2.14-2.13 (m, 2H).

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2016/115090, 2016, A1 Location in patent: Paragraph 0197-0204
[2]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0156-0163

73
[ 14441-90-8 ]
[ 924-73-2 ]
ethyl 3-(5-phenylisoxazole-3-carboxamido)propanoate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
53%	With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; triethylamine In tetrahydrofuran at 20℃; for 16h; Cooling with ice;	15.1 Step 1: ethyl 3-(5-phenylisoxazole-3-carboxamido)propanoate Step 1: ethyl 3-(5-phenylisoxazole-3-carboxamido)propanoate HOBt (7.50 g, 0.055 mol), ethyl 3-aminopropanoate (5.0 g, 42.0 mmol), triethylamine (17.30 g, 170 mmol) and EDC.HCl (12.20 g, 65.0 mmol) were added sequentially to an ice cooled stirred solution of 5-phenylisoxazole-3-carboxylic acid (8.07 g, 42.0 mmol) in THF (170 mL) and the reaction mixture was stirred at room temperature for 16 h. Volatiles were concentrated under reduced pressure and ice-water (200 mL) was added to the crude reaction mixture. The precipitate thus obtained was filtered and dried to afford crude compound which was further purified by flash column chromatography using 25% EtOAc in hexane as eluent to obtain the product (6.5 g, 53%) as off-white solid. 1H NMR (400 MHz, CDCl3): δ 7.79-7.76 (m, 2H), 7.50-7.45 (m, 3H), 7.35 (br s, 1H), 6.94 (s, 1H), 4.17 (q, J=7.1 Hz, 2H), 3.75-3.71 (m, 2H), 2.66-2.63 (t, J=6.1 Hz, 2H), 1.27 (t, J=7.1 Hz, 3H); LC-MS: [M+H]+ 289.1.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - US2017/1993, 2017, A1 Location in patent: Paragraph 0232

74
ethyl 5-(2-aminoethyl)-1,3,4-thiadiazole-2-carboxylate [ No CAS ]
[ 14441-90-8 ]
ethyl 5-(2-(5-phenylisoxazole-3-carboxamido)ethyl)-1,3,4-thiadiazole-2-carboxylate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
48.2%	With N-ethyl-N,N-diisopropylamine; HATU In tetrahydrofuran at 20℃; for 4h;	9.1 Step 1: ethyl 5-(2-(5-phenylisoxazole-3-carboxamido)ethyl)-1,3,4-thiadiazole-2-carboxylate Step 1: ethyl 5-(2-(5-phenylisoxazole-3-carboxamido)ethyl)-1,3,4-thiadiazole-2-carboxylate HATU (0.45 g, 0.001 mol) was added to a solution of 5-phenylisoxazole-3-carboxylic acid (0.15 g, 0.7 mmol) and ethyl 5-(2-aminoethyl)-1,3,4-thiadiazole-2-carboxylate (0.2 g, 1 mmol) in THF (4 mL) followed by DIPEA (0.3 g, 2 mmol) and the resulting reaction mixture was stirred at room temperature for 4 h. Progress of the reaction was monitored by TLC. The reaction mixture was poured onto ice cooled water (10 mL) and the precipitate thus formed was filtered. Residue was washed with water and hexane (2*10 mL) and dried under reduced pressure to afford the product (0.14 g, 48.2%) as off white solid. 1H NMR (400 MHz, CDCl3): δ 7.79-7.76 (m, 2H), 7.50-7.45 (m, 3H), 7.38 (t, J=5.6 Hz, 1H), 6.93 (s, 1H), 4.50 (q, J=7.1 Hz, 2H), 4.00 (q, J=6.3 Hz, 2H), 3.51 (t, J=6.4 Hz, 2H), 1.44 (t, J=7.1 Hz, 3H); LC-MS: [M+H]+=373.7.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - US2017/1993, 2017, A1 Location in patent: Paragraph 0176

75
[ 14441-90-8 ]
2-(5-methoxymethyl-[1,3,4]thiadiazol-2-yl)ethylamine [ No CAS ]
N-(2-(5-(methoxymethyl)-1,3,4-thiadiazol-2-yl)ethyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
48%	With 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane-2,4,6-trioxide; triethylamine In dichloromethane; ethyl acetate at 20℃; for 18h; Cooling with ice;	2.4 Step-4: N-(2-(5-(methoxymethyl)-1,3,4-thiadiazol-2-yl)ethyl)-5-phenylisoxazole-3-carboxamide Step-4: N-(2-(5-(methoxymethyl)-1,3,4-thiadiazol-2-yl)ethyl)-5-phenylisoxazole-3-carboxamide To an ice-cooled suspension of 5-phenyl-isoxazole-3-carboxylic acid (2.08 g, 0.011 mol) in DCM (40 mL) was slowly added T3P solution (10.5 mL, 0.016 mol, 50% solution in EtOAc) followed by addition of triethylamine (2.23 g, 0.022 mol) and solution of 2-(5-methoxymethyl-[1,3,4]thiadiazol-2-yl)-ethylamine (1.88 g, ca. 0.011 mol) in DCM (10 mL). The reaction mixture was stirred at room temperature for 18 h and quenched onto ice-water (150 mL). Separated DCM layer was washed with saturated NaHCO3 solution (375 mL), water (250 mL) and brine (100 mL). Organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum to obtained crude solid product. The crude solid was triturated in diethyl ether (50 mL), filtered and dried to afford desired product. Yield: 48%; Appearance: off-whites solid Analytical data: 1H NMR (400 MHz, CDCl3): δ 7.79-7.76 (m, 2H), 7.50-7.46 (m, 3H), 7.41 (br, 1H), 6.93 (s, 1H), 4.82 (s, 2H), 3.99-3.94 (m, 2H), 3.45 (s, 3H), 3.42-3.41 (m, 2H). LC-MS: (M+H)+ 345.0; HPLC Purity: 99.87% at 254 nm & 99.32% at 220 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - US2017/1993, 2017, A1 Location in patent: Paragraph 0134

76
[ 14441-90-8 ]
2-(5-methoxymethyl-[1,3,4]oxadiazol-2-yl)ethylamine [ No CAS ]
N-(2-(5-(methoxymethyl)-1,3,4-oxadiazol-2-yl)ethyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
48%	With 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane-2,4,6-trioxide; triethylamine In tetrahydrofuran at 20℃; for 12h;	5.3 Step-3: N-(2-(5-(methoxymethyl)-1,3,4-oxadiazol-2-yl)ethyl)-5-phenylisoxazole-3-carboxamide Step-3: N-(2-(5-(methoxymethyl)-1,3,4-oxadiazol-2-yl)ethyl)-5-phenylisoxazole-3-carboxamide A solution of 2-(5-methoxymethyl-[1,3,4]oxadiazol-2-yl)-ethylamine (100 mg, 0.00064 mol), 5-phenylisoxazole-3-carboxylic acid (144 mg, 0.00076 mol), TEA (0.13 ml, 0.00093 mol) and T3P (0.303 mg, 0.00093 mol) in THF (5 mL) was stirred at room temperature for 12 h at room temperature. The reaction mixture was evaporated, diluted with aqueous NaHCO3 solution (5 mL) and extracted with EtOAc (2*5 ml). Combined organic layers were washed with water, brine, dried over sodium sulfate and concentrated under vacuum to afford crude product. The crude compound was purified by preparative HPLC to obtain compound (100 mg) as off-white solid. Yield: 48% Appearance: off-white solid Analytical data: 1H NMR (400 MHz, CDCl3): δ 7.79-7.76 (m, 2H), 7.49-7.45 (m, 3H), 7.40 (br, 1H), 6.93 (s, 1H), 4.62 (s, 2H), 3.98-3.94 (q, 2H, J=6.4 Hz), 3.45 (s, 3H), 3.21-3.18 (t, 2H, J=4.7 Hz). (0152) LC-MS: [M+H]+=329.2; HPLC purity: 98.26% at 220 nm, 99.23% at 254 nm.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - US2017/1993, 2017, A1 Location in patent: Paragraph 0149

77
[ 14441-90-8 ]
trans-3-(1H-imidazol-1-yl)cyclobutan-1-amine hydrochloride [ No CAS ]
5-phenyl-N-[trans-3-(1H-imidazol-1-yl)cyclobutyl]isoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
6%	With N-ethyl-N,N-diisopropylamine; HATU In dichloromethane at 20℃; for 2h;	62.4 5-phenyl-N-[trans-3-(1H-imidazol-1-yl)cyclobutyl]isoxazole-3-carboxamide fram,-3-(lH-inridazol-l-yl)cyclobutan-l-amine hydrochloride (400 mg, 2.30 mmol, 0.70 eq.), HATU (1.507 g, 3.96 mmol, 1.20 eq.) and DIEA (1.279 g, 9.90 mmol, 3.00 eq.) were added to a solution of 5-phenylisoxazole-3-carboxylic acid (624 mg, 3.30 mmol, 1.00 eq.) in dichloromethane (30 mL). The resulting solution was stirred for 2 hours at room temperature and it was then concentrated under vacuum. The crude product was purified by Prep-HPLC with the following conditions (Waters): Column, XBridge BEH130 Prep C18 OBD Column, 19* 150mm 5um 13nm; mobile phase, water with 0.05%TFA and ACN (12.0% ACN up to 50.0% in 8 min); Detector, UV 254nm. This resulted in 58 mg (6%) of 5-phenyl-N- [fra ,-3-(lH-imidazol-l-yl)cyclobutyl]isoxazole-3-carboxamide as a white solid. [0579] Analytical data: [0580] LC-MS (ES, m/z): [M+H]+ = 309.2 [0581] HPLC purity: 99.4% at 254 nm [0582] NMR (300MHz, OMSO-d6, ppm): δ 9.28 (s, 1H), 7.98-7.93 (m, 3H), 7.51 (s, 1H), 7.60-7.56 (m, 3H), 7.37 (s, 1H), 5.20-5.11 (m, 1H), 4.69-4.60 (m, 1H), 2.91-2.45 (m, 4H).

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - WO2017/40606, 2017, A1 Location in patent: Paragraph 0578-0582

78
[ 14441-90-8 ]
1-isopropyl-1H-pyrazolo [3, 4-d] pyrimidine-3, 4-diamine [ No CAS ]
N-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
53%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With oxalyl dichloride; N,N-dimethyl-formamide In dichloromethane at 0℃; Inert atmosphere; Stage #2: 1-isopropyl-1H-pyrazolo [3, 4-d] pyrimidine-3, 4-diamine With N-ethyl-N,N-diisopropylamine In tetrahydrofuran; dichloromethane at -10℃; for 3h; Inert atmosphere;	1.9 General procedure for the synthesis of compounds 21a-o General procedure: To a solution of aryl or alkyl acid (1 equiv) in CH2Cl2 (10 mL), was added oxalyl chloride (2.5 equiv) and DMF (0.01 equiv) at 0°C and then stirring for 2h. To the resulting solid were added THF (10 mL) and 20 (1.2 equiv) after concentrating under reduced pressure, subsequent cooling to -10°C, DIPEA (2.5 equiv) was added dropwise, and the reaction proceed for 3 h. The mixture was added H2O and extracted with chloroform. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (MeOH/DCM=30/1) to afford the corresponding product.

Reference： [1]Wang, Chengyan; Liu, Hongchun; Song, Zilan; Ji, Yinchun; Xing, Li; Peng, Xia; Wang, Xisheng; Ai, Jing; Geng, Meiyu; Zhang, Ao [Bioorganic and Medicinal Chemistry Letters, 2017, vol. 27, # 11, p. 2544 - 2548]

79
[ 14441-90-8 ]
[ 1354355-91-1 ]
N-((2',3-dimethyl-[2,4'-bipyridin]-5-yl)methyl)-5-phenylisoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
79.8%	With N-ethyl-N,N-diisopropylamine; HATU In dimethyl sulfoxide at 20℃; for 2h;	6.2 Step2: Preparation of Compound REX-P-7 To a solution of 1-2 (0.7 g, 3.9 mmol) and REX-P-INT-1 (0.9 g, 4.3 mmol) in DMSO (10 mL) was added N,N-diisopropylethylamine (1.6 mL, 11.7 mmol) and o-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (1.9 g, 5.0 mmol). The reaction mixture was stirred at room temperature for 2 hours and quenched with ice water (200 mL), extracted with EtOAc, washed with brine, dried with anhydrous Na2SO4, filtered and concentrated to give the crude product. The crude product was purified by silica gel chromatography eluted to give product REX-P-7 (1.2 g, 79.8%). MS m/z [ESI]: 385.2 [M+1]. 1H-NMR (400 MHz, DMSO-d6): δ=9.47 (t, J=6.0 Hz, 1H), 8.48-8.65 (m, 2H), 7.90-7.98 (m, 2H), 7.71 (s, 1H), 7.52-7.61 (m, 3H), 7.37-7.43 (m, 2H), 7.34 (d, J=4.8 Hz, 1H), 4.54 (d, J=6.0 Hz, 2H), 2.54 (s, 3H), 2.34 (s, 3H).

Reference： [1]Current Patent Assignee: SUZHOU SINOVENT PHARMACEUTICALS - US2018/271846, 2018, A1 Location in patent: Paragraph 0159; 0163-0165

80
[ 92-54-6 ]
[ 14441-90-8 ]
(5-phenyl-1,2-oxazol-3-yl)(4-phenylpiperazin-1-yl)methanone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
74%	With O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 25℃; for 2h;	133 Example 133. Example 133. Preparation of 5-phenylisoxazol-3-yl)(4-phenylpiperazin-1-yl)methanone (266) To a stirred solution of 5-phenylisoxazole-3-carboxylic acid (0.150 g, 0.793 mmol) and N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (0.301 g, 0.793 mmol) in N,N'-dimethylformamide (2 mL) was added diisopropylethylamine (0.205 g, 1.59 mmol) and 1-phenylpiperazine (0.129 g, 0.793 mmol). The mixture was stirred at 25° C. for 2 h then purified directly by prep-HPLC (Luna C18 100*30 5 μm column; 53-83% acetonitrile in a 10 mM ammonium bicarbonate solution in water, 10 min gradient) to give ((5-phenylisoxazol-3-yl) (4-phenylpiperazin-1-yl)methanone (0.198 g, 0.589 mmol, 74%) as a white solid. 1H NMR (400 MHz, Dimethylsulfoxide-d6) δ 7.94 (dd, J=1.5, 7.7 Hz, 2H), 7.60-7.52 (m, 3H), 7.33 (s, 1H), 7.27-7.20 (m, 2H), 6.97 (d, J=7.9 Hz, 2H), 6.82 (t, J=7.2 Hz, 1H), 3.81 (td, J=4.9, 15.1 Hz, 4H), 3.26-3.15 (m, 4H); LCMS (ESI) m/z: 334.2 [M+H]+.
	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In acetonitrile at 20℃; for 1h; Stage #2: 4-phenyl-1-piperazine In acetonitrile at 20℃;

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - US2019/330198, 2019, A1 Location in patent: Paragraph 1370
[2]Saeedi, Mina; Mohtadi-Haghighi, Dorrin; Mirfazli, Seyedeh Sara; Mahdavi, Mohammad; Hariri, Roshanak; Lotfian, Hania; Edraki, Najmeh; Iraji, Aida; Firuzi, Omidreza; Akbarzadeh, Tahmineh [Chemistry and biodiversity, 2019, vol. 16, # 2]

81
[ 14441-90-8 ]
1-((4-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazol-4-amine [ No CAS ]
5-phenyl-N-(1-((4-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazol-4-yl)isoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
25%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With oxalyl dichloride In dichloromethane at 20℃; for 0.5h; Stage #2: 1-((4-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazol-4-amine With triethylamine In dichloromethane at 20℃; for 0.5h;	55.2 Step 2: Preparation of 5-phenyl-N-(1-((4-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazol-4-yl)isoxazole-3-carboxamide To a solution of 5-phenylisoxazole-3-carboxylic acid (117 mg, 0.620 mmol) in dichloromethane (2 mL) at 20° C. was added oxalyl chloride (1 mL). The reaction mixture was stirred at room temperature for 0.5 h. The volatiles were removed in vacuo. The crude residue was dissolved in dichloromethane (2 mL) and added to a mixture of 1-((4-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazol-4-amine (0.150 g, 0.620 mmol) and triethylamine (0.188 g, 1.86 mmol) in dichloromethane (5 mL) dropwise. The reaction mixture was stirred for 0.5 h and purified by prep-HPLC (the crude sample was dissolved in N,N-dimethylformamide and loaded onto Boston C18 21×250 mm 10 μm column. The mobile phases were acetonitrile/0.01% aqueous trifluoroacetic acid) to offer 5-phenyl-N-(1-((4-(trifluoromethyl)pyridin-3-yl)methyl)-1H-pyrazol-4-yl)isoxazole-3-carboxamide (65.5 mg, 0.158 mmol, 25%) as a white solid. 1H NMR (500 MHz, Dimethylsulfoxide-d6) δ 11.09 (s, 1H), 8.81 (d, J=5.0 Hz, 1H), 8.34 (s, 1H), 8.29 (s, 1H), 7.98 (dd, J=7.6, 1.6 Hz, 2H), 7.80 (d, J=5.0 Hz, 1H), 7.75 (s, 1H), 7.59-7.56 (m, 3H), 7.47 (s, 1H), 5.61 (s, 2H); LCMS (ESI) m/z: 414.1 [M+H]+.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - US2019/330198, 2019, A1 Location in patent: Paragraph 0976

82
[ 14441-90-8 ]
[ 28466-62-8 ]
N-(1-benzylpyrazol-4-yl)-5-phenyl-isoxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
36%	With O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 15℃; for 3h;	Step 3: Preparation of N-(1-benzylpyrazol-4-yl)-5-phenyl-isoxazole-3-carboxamide To a stirred solution of <strong>[28466-62-8]1-benzylpyrazol-4-amine</strong> (0.120 g, 0.693 mmol) and 5-phenylisoxazole-3-carboxylic acid (0.157 g, 0.831 mmol) in N,N-dimethylformamide (1 mL) was added N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (0.315 g, 0.831 mmol) and diisopropylethylamine (0.242 mL, 1.39 mmol) at 15 C., then stirred at 15 C. for 3 h. The reaction was purified by prep-HPLC (Agela Venusil XBP C18 150*25 5 um column; 50%-75% acetonitrile in a 10 mM 0.04% ammonium hydroxide, 10 min gradient) to give N-(1-benzylpyrazol-4-yl)-5-phenyl-isoxazole-3-carboxamide (0.087 g, 0.253 mmol, 36%) as a pale yellow solid. 1H NMR (400 MHz, Chloroform-d) delta 8.57 (br. s, 1H), 8.05 (s, 1H), 7.86-7.82 (m, 2H), 7.65 (s, 1H), 7.54-7.50 (m, 3H), 7.41-7.33 (m, 3H), 7.30-7.27 (m, 2H), 7.03 (s, 1H), 5.32 (s, 2H); LCMS (ESI) m/z: 345.1 [M+H]+.

Reference： [1]Patent: US2019/330198,2019,A1 .Location in patent: Paragraph 0587

83
[ 308823-90-7 ]
[ 14441-90-8 ]
(5-phenylisoxazol-3-yl)-[4-[2-(trifluoromethyl)phenyl]-1-piperidyl]methanone [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
64%	With 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane-2,4,6-trioxide; triethylamine In dichloromethane; ethyl acetate at 25℃; for 15h;	114 Example 114. Example 114. Preparation of 5-phenylisoxazol-3-yl)-[4-[2-(trifluoromethyl)phenyl]-1-piperidyl]methanone (289) To a stirred solution of 5-phenylisoxazole-3-carboxylic acid (0.100 g, 0.529 mmol) in dichloromethane (3 mL) was added 4-[2-(trifluoromethyl)phenyl]piperidine (0.155 g, 0.582 mmol), triethylamine (0.160 g, 1.59 mmol) and propylphosphonic anhydride (0.505 g, 0.793 mmol, 50% in ethyl acetate). The mixture was stirred at 25° C. for 15 h. The reaction mixture was quenched with water (10 mL) and extracted with dichloromethane (10 mL3). The combined organic layers were washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (YMC-Actus Triart C18 10030 mm*5 μm column; 60-90% acetonitrile in a 10 mM ammonium bicarbonate solution in water, 12 min gradient) to offer (5-phenylisoxazol-3-yl)-[4-[2-(trifluoromethyl)phenyl]-1-piperidyl]methanone (0.135 g, 0.336 mmol, 64%) as a pale yellow solid. 1H NMR (400 MHz, Chloroform-d) δ 7.82 (dd, J=1.8, 7.5 Hz, 2H), 7.66 (d, J=7.9 Hz, 1H), 7.57-7.43 (m, 5H), 7.33 (t, J=7.7 Hz, 1H), 6.85 (s, 1H), 4.95 (br. d, J=13.6 Hz, 1H), 4.69 (br. d, J=13.6 Hz, 1H), 3.35-3.21 (m, 2H), 2.92 (dt, J=2.9, 13.0 Hz, 1H), 2.02-1.75 (m, 4H); LCMS (ESI) m/z: 401.1 [M+H]+.

Reference： [1]Current Patent Assignee: YUMANITY THERAPEUTICS INC - US2019/330198, 2019, A1 Location in patent: Paragraph 1308

84
[ 14441-90-8 ]
[ 118979-65-0 ]
N-[3-(azepan-1-yl)propyl]-5-phenyl-1,2-oxazole-3-carboxamide [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
28%	With N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 20℃;	Synthesis of AIM-0067, AIM-0071 and AIM-0078 (general combinatorial procedure) General procedure: To a solution of HATU (61 mg, 0.16 mmol) and DIPEA (28 μL, 0.16 mmol) in DMF (0.50 mL) was addedcorresponding carboxylic acid (160 μmol). 3-(Azepan-1-yl)propan-1-amine dihydrochloride (18 mg, 80 μmol) andDIPEA (28 μL, 0.16 mmol) in DMF (0.50 mL) were added to the reaction mixture. The mixture was stirred at roomtemperature overnight. The mixture was poured into water at room temperature and evaporated by blowing awaywith the air at 60 °C. The residue was purified by preparative HPLC (YMCTriartC18, eluted with MeCN/10mMNH4HCO3 aq.). The desired fraction was concentrated by blowing away with the air at 60 °C to give the product.

Reference： [1]Kabeche, Stephanie; Aida, Jumpei; Akther, Thamina; Ichikawa, Takashi; Ochida, Atsuko; Pulkoski-Gross, Michael J.; Smith, Mark; Humphries, Paul S.; Yeh, Ellen [Bioorganic and Medicinal Chemistry Letters, 2021, vol. 41]

85
[ 32418-24-9 ]
[ 14441-90-8 ]
ethyl 2-((5-(5-phenylisoxazole-3-carboxamido)-1,3,4-thiadiazol-2-yl)thio)acetate [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
52%	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In acetonitrile at 20℃; for 1h; Stage #2: ethyl 2-[(5-amino-1,3,4-thiadiazol-2-yl)thio]acetate In acetonitrile	2.2.1. General Procedure for the Preparation of 5-arylisoxazole-1,3,4-thiadiazole 5 General procedure: The mixture of 5-arylisoxazole-3- carboxylic acid derivative2 (1 mmol), hydroxybenzotriazole (HOBt, 1 mmol) andN-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride(EDCI, 1 mmol) was stirred in dry acetonitrile atroom temperature for 1 h; then ethyl 2-((5-amino-1,3,4-thiadiazol-2-yl)thio)acetate (4) (1 mmol) was added to themixture and the reaction was continued for 24-72 h. Aftercompletion of the reaction (checked by TLC), the solventwas evaporated under vacuum, and the crude was extractedusing chloroform and washed with water and solutions ofNaHCO3 10%, saturated NaCl, and citric acid, respectively.After drying the organic phase over sodium sulfate, the solventwas removed and the solid product was recrystallizedfrom ethyl acetate and petroleum ether. 2.2.1.1. Ethyl 2-((5-(5-phenylisoxazole-3-carboxamido)-1,3,4-thiadiazol-2-yl)thio)acetate (5a)Yield: 52%; M.p. = 205-207 °C; IR (KBr, cm-1): 3490,3214, 3099, 2989, 1734, 1691, 1644, 1618, 1539, 1448. 1HNMR (500 MHz, DMSO-d6): δ = 13.75 (s,1H, NH), 7.95 (d,J= 7.1 Hz, 2H, H2, H6), 7.61-7.57 (m, 4H, H3, H4, H5, isoxazole),4.21 (s, 2H, CH2), 4.14 (q, J = 7.0 Hz, 2H, OCH2),1.19 (t, J = 7.0 Hz, 3H, CH3) ppm. 13C NMR (125 MHz, DMSO- d6): δ = 170.9, 168.2, 162.7, 158.2, 157.7, 155.2,131.1, 129.4, 126.0, 125.8, 100.3, 61.4, 35.0, 14.0 ppm.Calcd for C16H14N4O4S2: C, 49.22; H, 3.61; N, 14.35. Found:C, 49.54; H, 3.80; N, 14.58.

Reference： [1]Saeedi, Mina; Eslami, Azadeh; Mirfazli, Seyedeh Sara; Zardkanlou, Mahsa; Faramarzi, Mohammad Ali; Mahdavi, Mohammad; Akbarzadeh, Tahmineh [Letters in drug design and discovery, 2021, vol. 18, # 5, p. 436 - 444]

86
[ 14441-90-8 ]
[ 37561-42-5 ]
C₂₂H₂₂N₂O₄ [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; N-[3-(N,N-dimethylamino)-propyl]-N'-ethyl-carbodiimide hydrochloride In N,N-dimethyl-formamide at 20℃; for 0.5h; Stage #2: 2,2-dimethyl-5-phenyl-1,3-dioxan-5-amine With N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 5h;	5.15. General procedure for the synthesis of intermediates 41a-al General procedure: The aryl acid (1.0 equiv.), HOBt (1.1 equiv.) and EDCI (1.1 equiv.)were added to DMF, and the mixture was stirred at room temperaturefor 30 min. Then amino 39a-g and DIEA (1.1 equiv.) wereadded to the reaction and continued to stir for 5 h. The reactionwaspoured in water and the mixture was extracted with ethyl acetatethree times. The combined extracts were dried and concentratedunder reduced pressure to afford 41a-al.

Reference： [1]Cheng, Maosheng; Su, Xin; Sun, Nannan; Sun, Yin; Tian, Linfeng; Yin, Wenbo; Zhang, Chu; Zhao, Dongmei; Zhao, Liyu; Zhao, Shizhen; Zheng, Yang [European Journal of Medicinal Chemistry, 2022, vol. 228]

87
[ 14441-90-8 ]
[ 5680-80-8 ]
C₁₄H₁₄N₂O₅*ClH [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; N-[3-(N,N-dimethylamino)-propyl]-N'-ethyl-carbodiimide hydrochloride In N,N-dimethyl-formamide at 20℃; for 0.5h; Stage #2: (l)-serine methyl ester hydrochloride In N,N-dimethyl-formamide	4.2. General procedure for the synthesis of intermediates 10a-k General procedure: To a solution of different acids (2.45 mmol, 0.50 g) in DMF, HOBt(2.70 mmol, 0.36 g) and EDCI (2.70 mmol, 0.52 g) were added andthe mixture was stirred for 30 min at room temperature. Then, LSerineMethyl Ester Hydrochloride (2.70 mmol, 0.42 g) and DIEA(4.90 mmol, 0.64 g) were added. The reaction was stirred andmonitored with TLC. The mixture was poured into water andextracted with ethyl acetate three times. The combined organiclayers were dried over Na2SO4 and filtered. The filtrate was evaporatedunder reduced pressure to afford intermediates 10a-k, yield73.71% [17].
	Stage #1: 5-phenyl-3-isoxazolecarboxylic acid With benzotriazol-1-ol; N-[3-(N,N-dimethylamino)-propyl]-N'-ethyl-carbodiimide hydrochloride In N,N-dimethyl-formamide at 20℃; for 0.5h; Stage #2: (l)-serine methyl ester hydrochloride In N,N-dimethyl-formamide	4.2. General procedure for the synthesis of intermediates 10a-k General procedure: To a solution of different acids (2.45 mmol, 0.50 g) in DMF, HOBt(2.70 mmol, 0.36 g) and EDCI (2.70 mmol, 0.52 g) were added andthe mixture was stirred for 30 min at room temperature. Then, LSerineMethyl Ester Hydrochloride (2.70 mmol, 0.42 g) and DIEA(4.90 mmol, 0.64 g) were added. The reaction was stirred andmonitored with TLC. The mixture was poured into water andextracted with ethyl acetate three times. The combined organiclayers were dried over Na2SO4 and filtered. The filtrate was evaporatedunder reduced pressure to afford intermediates 10a-k, yield73.71% [17].

Reference： [1]Cheng, Maosheng; Cui, Hengxian; Gao, Zixuan; Jiang, Hong; Liu, Lei; Qin, Qiaohua; Su, Xin; Sun, Yin; Wu, Tianxiao; Yin, Wenbo; Zhang, Yuxin; Zhao, Dongmei; Zhao, Liyu [European Journal of Medicinal Chemistry, 2022, vol. 233]
[2]Cheng, Maosheng; Cui, Hengxian; Gao, Zixuan; Jiang, Hong; Liu, Lei; Qin, Qiaohua; Su, Xin; Sun, Yin; Wu, Tianxiao; Yin, Wenbo; Zhang, Yuxin; Zhao, Dongmei; Zhao, Liyu [European Journal of Medicinal Chemistry, 2022, vol. 233]

88
[ 14441-90-8 ]
[ 1446-61-3 ]
C₃₀H₃₆N₂O₂ [ No CAS ]

Yield	Reaction Conditions	Operation in experiment
90%	With 4-dimethylaminopyridine; N-[3-(N,N-dimethylamino)-propyl]-N'-ethyl-carbodiimide hydrochloride In dichloromethane at 25℃;	29 Example 29 Take 5-phenyl-3-isoxazolecarboxylic acid (1.20mmol) and dehydroabietylamine (1.0mmol) in a single-necked flask, add dichloromethane (10mL) to dissolve, add catalyst 4-dimethylaminopyridine ( 0.20 mmol) and condensing agent EDCI (1.20 mmol), added, reacted at 25°C for 6-10 h, detected by TLC, the reaction was completed, the organic layer was washed with saturated sodium bicarbonate solution, washed with water, washed with saturated brine, and anhydrous sodium sulfate Dry, filter with suction, and concentrate under reduced pressure to remove dichloromethane to obtain crude product, which is purified by silica gel column chromatography (eluent dichloromethane) and dried to obtain target compound I-29. Yield 90%, white solid, mp240-242.5°C.

Reference： [1]Current Patent Assignee: NANJING FORESTRY UNIVERSITY - CN113773288, 2021, A Location in patent: Paragraph 0100-0102

Purity	Size	Price	VIP Price	USA Stock *0-1 Day	Global Stock *5-7 Days	Quantity
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CAS No. :	14441-90-8	MDL No. :	MFCD00464221
Formula :	C₁₀H₇NO₃	Boiling Point :	-
Linear Structure Formula :	-	InChI Key :	XJYOBHXWBRKOQO-UHFFFAOYSA-N
M.W :	189.17	Pubchem ID :	151916
Synonyms :

Num. heavy atoms :	14
Num. arom. heavy atoms :	11
Fraction Csp3 :	0.0
Num. rotatable bonds :	2
Num. H-bond acceptors :	4.0
Num. H-bond donors :	1.0
Molar Refractivity :	48.9
TPSA :	63.33 Å²

GI absorption :	High
BBB permeant :	Yes
P-gp substrate :	No
CYP1A2 inhibitor :	Yes
CYP2C19 inhibitor :	No
CYP2C9 inhibitor :	No
CYP2D6 inhibitor :	No
CYP3A4 inhibitor :	No
Log Kp (skin permeation) :	-5.73 cm/s

Log Po/w (iLOGP) :	0.96
Log Po/w (XLOGP3) :	2.43
Log Po/w (WLOGP) :	2.04
Log Po/w (MLOGP) :	1.03
Log Po/w (SILICOS-IT) :	1.78
Consensus Log Po/w :	1.65

[ CAS No. 14441-90-8 ] {[proInfo.proName]}

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[ 14441-90-8 ] Synthesis Path-Upstream 1~4

[ 14441-90-8 ] Synthesis Path-Downstream 1~88

Related Functional Groups of
[ 14441-90-8 ]

Aryls

Carboxylic Acids

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[ 14441-90-8 ]

Isoxazoles

Precautionary Statements-General

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Inquiry

Lipinski :	0.0
Ghose :	None
Veber :	0.0
Egan :	0.0
Muegge :	1.0
Bioavailability Score :	0.56

Log S (ESOL) :	-2.99
Solubility :	0.192 mg/ml ; 0.00102 mol/l
Class :	Soluble
Log S (Ali) :	-3.4
Solubility :	0.0748 mg/ml ; 0.000396 mol/l
Class :	Soluble
Log S (SILICOS-IT) :	-3.13
Solubility :	0.14 mg/ml ; 0.00074 mol/l
Class :	Soluble

PAINS :	0.0 alert
Brenk :	0.0 alert
Leadlikeness :	1.0
Synthetic accessibility :	2.6

Signal Word:	Warning	Class:	N/A
Precautionary Statements:	P261-P280-P305+P351+P338	UN#:	N/A
Hazard Statements:	H302-H315-H319-H332-H335	Packing Group:	N/A
GHS Pictogram:

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Code	Phrase
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P221	Take any precaution to avoid mixing with combustibles
P222	Do not allow contact with air.
P223	Keep away from any possible contact with water, because of violent reaction and possible flash fire.
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P231	Handle under inert gas.
P232	Protect from moisture.
P233	Keep container tightly closed.
P234	Keep only in original container.
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P351	Rinse cautiously with water for several minutes.
P352	Wash with plenty of soap and water.
P353	Rinse skin with water/shower.
P360	Rinse immediately contaminated clothing and skin with plenty of water before removing clothes.
P361	Remove/Take off immediately all contaminated clothing.
P362	Take off contaminated clothing and wash before reuse.
P363	Wash contaminated clothing before reuse.
P370	In case of fire:
P371	In case of major fire and large quantities:
P372	Explosion risk in case of fire.
P373	DO NOT fight fire when fire reaches explosives.
P374	Fight fire with normal precautions from a reasonable distance.
P376	Stop leak if safe to do so. Oxidising gases (section 2.4) 1
P377	Leaking gas fire: Do not extinguish, unless leak can be stopped safely.
P378
P380	Evacuate area.
P381	Eliminate all ignition sources if safe to do so.
P390	Absorb spillage to prevent material damage.
P391	Collect spillage. Hazardous to the aquatic environment
P301 + P310	IF SWALLOWED: Immediately call a POISON CENTER or doctor/physician.
P301 + P312	IF SWALLOWED: call a POISON CENTER or doctor/physician IF you feel unwell.
P301 + P330 + P331	IF SWALLOWED: Rinse mouth. Do NOT induce vomiting.
P302 + P334	IF ON SKIN: Immerse in cool water/wrap in wet bandages.
P302 + P350	IF ON SKIN: Gently wash with plenty of soap and water.
P303 + P361 + P353	IF ON SKIN (or hair): Remove/Take off Immediately all contaminated clothing. Rinse SKIN with water/shower.
P304 + P312	IF INHALED: Call a POISON CENTER or doctor/physician if you feel unwell.
P304 + P340	IF INHALED: Remove victim to fresh air and Keep at rest in a position comfortable for breathing.
P304 + P341	IF INHALED: If breathing is difficult, remove victim to fresh air and keep at rest in a position comfortable for breathing.
P305 + P351 + P338	IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
P306 + P360	IF ON CLOTHING: Rinse Immediately contaminated CLOTHING and SKIN with plenty of water before removing clothes.
P307 + P311	IF exposed: call a POISON CENTER or doctor/physician.
P308 + P313	IF exposed or concerned: Get medical advice/attention.
P309 + P311	IF exposed or if you feel unwell: call a POISON CENTER or doctor/physician.
P332 + P313	IF SKIN irritation occurs: Get medical advice/attention.
P333 + P313	IF SKIN irritation or rash occurs: Get medical advice/attention.
P335 + P334	Brush off loose particles from skin. Immerse in cool water/wrap in wet bandages.
P337 + P313	IF eye irritation persists: Get medical advice/attention.
P342 + P311	IF experiencing respiratory symptoms: call a POISON CENTER or doctor/physician.
P370 + P376	In case of fire: Stop leak if safe to Do so.
P370 + P378	In case of fire:
P370 + P380	In case of fire: Evacuate area.
P370 + P380 + P375	In case of fire: Evacuate area. Fight fire remotely due to the risk of explosion.
P371 + P380 + P375	In case of major fire and large quantities: Evacuate area. Fight fire remotely due to the risk of explosion.
Storage
Code	Phrase
P401
P402	Store in a dry place.
P403	Store in a well-ventilated place.
P404	Store in a closed container.
P405	Store locked up.
P406	Store in corrosive resistant/ container with a resistant inner liner.
P407	Maintain air gap between stacks/pallets.
P410	Protect from sunlight.
P411
P412	Do not expose to temperatures exceeding 50 oC/ 122 oF.
P413
P420	Store away from other materials.
P422
P402 + P404	Store in a dry place. Store in a closed container.
P403 + P233	Store in a well-ventilated place. Keep container tightly closed.
P403 + P235	Store in a well-ventilated place. Keep cool.
P410 + P403	Protect from sunlight. Store in a well-ventilated place.
P410 + P412	Protect from sunlight. Do not expose to temperatures exceeding 50 oC/122oF.
P411 + P235	Keep cool.
Disposal
Code	Phrase
P501	Dispose of contents/container to ...
P502	Refer to manufacturer/supplier for information on recovery/recycling

[ CAS No. 14441-90-8 ] {[proInfo.proName]}

Quality Control of [ 14441-90-8 ]

Related Doc. of [ 14441-90-8 ]

Product Details of [ 14441-90-8 ]

Calculated chemistry of [ 14441-90-8 ]

Physicochemical Properties

Pharmacokinetics

Lipophilicity

Druglikeness

Water Solubility

Medicinal Chemistry

Safety of [ 14441-90-8 ]

Application In Synthesis of [ 14441-90-8 ]

[ 14441-90-8 ] Synthesis Path-Upstream 1~4

[ 14441-90-8 ] Synthesis Path-Downstream 1~88

Related Functional Groups of [ 14441-90-8 ]

Aryls

Carboxylic Acids

Related Parent Nucleus of [ 14441-90-8 ]

Isoxazoles

Precautionary Statements-General

Prevention

Response

Storage

Disposal

Physical hazards

Health hazards

Environmental hazards

Inquiry

Related Functional Groups of
[ 14441-90-8 ]

Related Parent Nucleus of
[ 14441-90-8 ]