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The BI-3802 was designed by Boehringer Ingelheim and could be obtained free of charge through the Boehringer Ingelheim open innovation portal opnMe.com, associated with its negative control.
L-Hydroxyproline is a versatile reagent for the synthesis of neuroexcitatory kainoids[1] and antifungal echinocandins.
Synonyms: L-Hydroxyproline; (2S,4R)-4-Hydroxypyrrolidine-2-carboxylic acid; trans-4-hydroxy L-Proline
4.5
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Martina Lioi ; Sara Tengattini ; Roberto Gotti ; Francesca Bagatin ; Stefano Galliani ; Gabriella Massolini , et al.
Abstract: During collagen biosynthesis, proline is post-translationally converted to hydroxyproline by specific enzymes. This amino acid, unique to collagen, plays a crucial role in stabilizing the collagen triple helix structure and could serve as an important biomarker for collagen content and quality analysis. Hydroxyproline has four isomers, depending on whether proline is hydroxylated at position 4 or 3 and on whether the cis- or trans- conformation is formed. Moreover, as extensive hydrolysis of collagen is required for its amino acid analysis, epimerization may also occur, although to a lesser extent, giving a total of eight possible isomers. The aim of the present study was to develop a reversed-phase high-performance liquid chromatography-UV-mass spectrometry (RPLC-UV-MS) method for the separation and quantification of all eight hydroxyproline isomers. After the chiral derivatization of the hydroxyproline isomers with Nα-(2,4-dinitro-5-fluorophenyl)-L-valinamide (L-FDVA), to enable their UV detection, the derivatized diastereoisomers were separated by testing different C18 column technologies and morphologies and optimizing operative conditions such as the mobile phase composition (solvent, additives), elution mode, flow rate and temperature. Baseline resolution of all eight isomers was achieved on a HALO® ES-C18 reversed-phase column (150×1.5 mm, 2.7 μm, 160 Å) using isocratic elution and MS-compatible mobile phase. The optimized method was validated for the quantification of hydroxyproline isomers and then applied to different collagen hydrolysates to gain insight and a deeper understanding of hydroxyproline abundances in different species (human, chicken) and sources (native, recombinant).
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Keywords: Collagen ; Amino acid analysis ; Hydroxyproline isomers ; Recombinant collagen ; Reverse phase chromatography ; Mass spectrometry
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Usman Sabir ; Hafiz Muhammad Irfan ; Alamgeer ; Aman Ullah ; Yusuf S. Althobaiti ; Fahad S. Alshehri , et al.
Abstract: Western diet style (fast food), which includes fatty frozen junk food, lard, processed meats, whole-fat dairy foods, cream, mayonnaise, butter, snacks, and fructose, is a primary etiological determinant for developing nonalcoholic steatohepatitis (NASH) worldwide. Here the primary focus is to see the impact of naturally identified essential oil on disease mechanisms developed in an animal model using the same ingredients. Currently, symptomatic therapies are recommended for the management of NASH due to non-availability of specific treatments. Therefore, the present study was designed to evaluate the potential anti-NASH effect of nerolidol in a rat model fed with a purpose-built diet. The diet substantially induced insulin resistance, hepatic steatosis, dyslipidemia, and elevation of liver enzymes in the experimental animals. The levels of liver oxidative stress markers, nitrites (NO2–), serum pro-inflammatory cytokine (TNF-α) and hepatic collagen were increased in disease control rats. Nerolidol oral treatment in ascending dose order of 250 and 500 mg/kg substantially reduced the steatosis (macrovesicular and microvesicular), degeneration of hepatocytes, and inflammatory cells infiltration. The amounts of circulatory TNF-α and tissue collagen were also reduced at 500 mg/kg dose of nerolidol, expressing its anti-fibrotic effect. The current study described the multiple-hit pathophysiology of NASH as enhanced steatosis, pro-inflammatory markers, and oxidative stress in rats, which resulted in the development of vicious insulin resistance. Nerolidol treatment significantly reduced hepatic lipid accumulation and halted disease progression induced by a hypercaloric diet.
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Keywords: Western diet ; Non-alcoholic steatohepatitis ; Insulin resistance ; Inflammation ; Nerolidol
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CAS No. : | 51-35-4 |
Formula : | C5H9NO3 |
M.W : | 131.13 |
SMILES Code : | O=C(O)[C@H]1NC[C@H](O)C1 |
Synonyms : |
L-Hydroxyproline; (2S,4R)-4-Hydroxypyrrolidine-2-carboxylic acid; trans-4-hydroxy L-Proline
|
MDL No. : | MFCD00064320 |
GHS Pictogram: |
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Signal Word: | Warning |
Hazard Statements: | H315-H319-H335 |
Precautionary Statements: | P261-P305+P351+P338 |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
93% | Example 4: Synthesis of (4R,5S,6S)- 3-[[(3S,5S)-5-[(dimethylamino)carbonyl]-3-ρyrrolidinyl]thio]-6-[(lR)-l-hydroxyethyl]-4-methyl- 7-oxo-l-azabicyclo[3,2,0]hept-2-ene-2-carboxylic acid (I)Synthesis of (2S,4S)-2-dimethylaminocarbonyl -4-thio-l-PNZ-pyrrolindine (XX)L-hydoxyproline (XXVI) (26.2 g, 0.20 mol) was added to a solution of sodium hydroxide (220 ml, 2N) and cooled to O0C to 50C. The solution of p-nitrobenzyl chloroformate dissolved in <n="17"/>methylene chloride (40 ml) was added dropwise. After stirred at the same temperature for 1 h, the phase of methylene chloride was separated. The aqueous phase was washed with methylene chloride (70 ml) and acidified with concentrated sulfuric acid (36.6 g) at a temperature of O0C to 5 C . Substantive crystals were precipitated, collected by filtration under vacuum, washed with water, dried, and afford trans- l-(p-nitrobenzyloxycarbonyl) -4-hydroxyl-L-proline (XXV) (57.8 g, yield 93%), m.p.: 134~135.5C. | |
93% | Example 6] Synthesis of (2S,4S)-2-dimethylaminocarbonyl -4-acetylthio-l-PNZ-pyrrolidine (XIX)1. Synthesis of trans- l-(p-nitrobenzyloxycarbonyl) -4-hydroxyl-L-proline (XV) L-hydoxyproline (XXVI) (26.2 g, 0.20 mol) was added to a solution of sodium hydroxide(220 ml, 2N) and cooled to 0C to 5C. The solution, of p-nitrobenzyl chloroformate dissolved in methylene chloride (40 ml) was added dropwise. After stirred at the same temperature for 1 h, the phase of methylene chloride was separated. The aqueous phase was washed with methylene chloride (70 ml) and acidified with concentrated sulfuric acid (36.6 g) at a temperature of 0C to 5 C . Substantive crystals were precipitated, collected by filtration under vacuum, washed with water, dried, and afford trans- l-(p-nitrobenzyloxycarbonyl) -4-hydroxyl-L-proline (XXV) (57.8 g, yield 93%), m.p.: 134~135.5C. | |
92.6% | With sodium hydroxide; In dichloromethane; water monomer; at 0 - 5℃; for 3h; | Add sodium hydroxide (66.9g, 1.68mol) and 600g of water to the reaction flask, after stirring and dissolving, add trans-L-hydroxyproline (100g, 0.76mol), stir and dissolve completely and then cool down to 0C, Add dropwise a solution of 50% p-nitrobenzyl chloroformate in dichloromethane (370 g, 0.86 mol), the temperature does not exceed 5 C; after the dropping is completed, the reaction is carried out at 0 to 5 C for 3 hours. The phases were extracted with 100 mL of dichloromethane; the aqueous phase was separated, and the pH was adjusted to 2-3 with 10% hydrochloric acid, then extracted three times with ethyl acetate (200 mL×3), and the organic phases were combined, washed with saturated brine, and washed with anhydrous sodium sulfate. It was dried, filtered, concentrated to dryness, and recrystallized with ethanol to obtain 219.2 g of white solid (intermediate 7) with a yield of 92.6%. |
80% | With sodium hydroxide; In water monomer; toluene; at 0 - 20℃; for 2h;Inert atmosphere; Schlenk technique; | trans-4-Hydroxy-L-proline (SI, 1.0 g, 7.6 mmol, 1.0 eq.) was dissolved in aqueous NaOH (0.5 M, 34 mL) and cooled in an ice bath. At 0 - 5 C, 4-nitrobenzoylchloroformate (1.9 g, 8.6 mmol, 1.1 eq.) in toluene (25 mL) was added dropwise. The bi-phasic mixture was stirred for two hours, after which the toluene (30 mL) was added and the phases separated. The aqueous phase was extracted with toluene and the combined organic phases extracted with aqueous NaOH (0.5 M). The aqueous phases were combined, adjusted to pH = 1 by adding cone. HCI and extracted with ethyl acetate (3 times). The extract was dried over sodium sulfate, filtered and concentrated in vacuo. Thus, compound 1 (1.89 g, 6.1 mmol, 80%) was obtained as off-white solid. ESI-LRMS for Ci3Hi5N207+[MH+]: calcd. 311.1 found 311.2 |
Trans-4-hydroxy-L-proline (II) was added to a solution of sodium hydroxide in water at 0-5 C, and stirred to get a clear solution, followed by p- nitrobenzyloxycarbonyl chloride in dichloromethane (MDC) was added and stirred till completion of reaction. at 0-5 C. To this reaction mass sodium hydroxide solution was added and the layers were separated. To the aqueous layer methanol was added and pH was adjusted to acidic using sulphuric acid at 0-5 C. The solid obtained was filtered, washed with water and dried in vacuum at 50C to afford the title compound (III) as colorless crystals. | ||
31.95 kg | With sodium hydroxide; In dichloromethane; water monomer; at 0 - 5℃; for 5h;Large scale; | 20L reactor water 12kg,1kg sodium hydroxide, stirred and dissolved, cooled to 25 ~ 30 ,Add L-hydroxyproline 15kg, stirring to dissolve,Cool to 0 ~ 5 ,The temperature was controlled at 0 ~ 5 2.7kg of p-nitrobenzyl chloroformate and 3kg of dichloromethane were added dropwise.Dropping about 2 hours, dropping is completed,Keep stirring 0 ~ 5 for about 3 hours. Detection reaction is complete.The dichloromethane phase was separated, the aqueous phase was washed with 3 kg of dichloromethane, the aqueous phase was separated,Control temperature but 15 degrees,Concentrated sulfuric acid adjusted to pH 2, cooled to 0 filtration,Washed, dried in the product(5) To a solution of (2S, 4R) -4- hydroxy- 1 - (((4-nitrobenzoyl) oxy) carbonyl) pyrrolidine-31.95kg |
With sodium hydroxide; In water monomer; toluene; at -5 - 0℃; for 1.25h;pH 9.0; | In a 500mL three-neck bottle added purified water 192g, add 15.3g of sodium hydroxide, stir and dissolve, and cool at 0 C; add 3g of 3-hydroxyproline, stir and dissolve; cool down at -5 C, added the drops of about 180g of p-nitrobenzyl chloroformate toluene solution, after about 1 hour, the drop is completed and pΗ=9; continue to stir for 15 minutes, and then let stand for 20 min, dispense, and obtained lower aqueous phase; in the lower aqueous phase added toluene 63mL X 3 and wash three times, stand still; control the internal temperature at 10 C, and add 6N hydrochloric acid to the aqueous phase, add dropwise until the water phase becomes turbid and stop adding, pH=4, add a small amount of seed crystals and stir until more crystals are precipitated, continue to add hydrochloric acid at pΗ = 2 (a total of about 5g of 6N dilute hydrochloric acid is required), cool down at 0 C, heat 1h suction filtration, wash with pure water to pΗ = 3, obtained compound I hydrate product. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
73.6% | (a) Add 400Kg L-hydroxyproline and 15Kg cyclohexenone toAfter stirring and dissolving in 1500Kg ethylene glycol monobutyl ether, the temperature is raised to 180,Incubate the reaction for 15 hours, after the detection reaction is complete, cool down and wait for treatment;(b) Extract the cooled solution in step (a) with 2000Kg of saturated brine in batches,After extraction, go directly to the next step.(c) The aqueous solution obtained in step (b), 800Kg sodium bicarbonate and 650KgBoc2O was added to 1000Kg 2-methyltetrahydrofuran, and stirring was continued for 15h at room temperature.After the test is complete,Filter the generated solid first, and then wash the solid 2-3 times with 200Kg methyl tert-butyl ether.The water phase was also washed 2-3 times with 600Kg methyl tert-butyl ether.(d) The organic phase obtained in step (c) is evaporated to dryness and added to 1200Kg petroleum ether,Then the temperature was lowered to -10C, a large amount of white solid was precipitated after stirring for 2h, filtered,Then use cold 300Kg petroleum ether to wash the solid, and after drying, the purity is 98.3% (R)-(-)-N-Boc-3-pyrrolidinol is a white solid of 386.7Kg, with a total yield of 73.6%. |
Tags: L-Hydroxyproline | Biologically Modified Amino Acid | Drug Standard | Pyrrolidines | Drug Analysis | Heterocyclic Building Blocks | Amino Acids | 51-35-4
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