| Size | Price | Stock | Qty |
|---|---|---|---|
| 1g |
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| Other Sizes |
| Targets |
Collagen biosynthesis; prolyl hydroxylase; collagen triple helix formation. cis-4-Hydroxy-L-proline is a proline analog that interferes with collagen triple helix formation by being incorporated into procollagen, producing non-helical procollagen that is retained in the ER and degraded intracellularly. Also inhibits Schwann cell differentiation and fibroblast growth.
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|---|---|
| ln Vitro |
In chick tendon fibroblasts, collagen production reduced from 25% to 7% of total protein synthesis. Cells degraded 8% of newly synthesized collagen normally; with cis-4-hydroxyproline, 25% of total collagen degraded to dialyzable peptides. Inhibits Schwann cell differentiation and accumulation of secreted collagen types I/IV and laminin. Suppresses growth of N-nitrosomethylurea-induced rat mammary tumors.
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| ln Vivo |
Inhibits fibroblast growth by preventing deposition of triple-helical collagen. Suppresses growth of N-nitrosomethylurea-induced rat mammary tumors. In animal models, causes intracellular accumulation of non-helical procollagen retained in rough ER, not passing to smooth ER or Golgi, leading to increased intracellular degradation of newly synthesized collagen.
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| Enzyme Assay |
(1) Prolyl hydroxylase inhibition: incubate purified prolyl hydroxylase (0.1-1 ug) with (Pro-Pro-Gly)10, 2-oxoglutarate (0.1 mM), Fe2+ (0.05 mM), ascorbate (1 mM), and cis-4-hydroxy-L-proline (0.1-1000 uM) in 50 mM Tris-HCl pH7.5 at 37degC for 30 min. (2) Measure hydroxylation by 14CO2 release from [1-14C]-2-oxoglutarate or by HPLC for hydroxyproline.
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| Cell Assay |
(1) Seed chick tendon fibroblasts or Schwann cells (5×10⁵/well) in DMEM+10% FBS. (2) Treat with cis-4-hydroxy-L-proline (0.1-10 mM) for 24-72 h. (3) For collagen synthesis: add 3H-proline (2 uCi/mL) for last 4-24 h. (4) Harvest cells and medium, precipitate procollagen with 30% ammonium sulfate, digest with pepsin, separate collagens by SDS-PAGE, detect by fluorography. (5) For intracellular degradation: after labeling, chase 0-24 h, measure TCA-soluble radioactivity. (6) Cell proliferation: treat 48-96 h, MTT/CCK-8.
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| Animal Protocol |
(1) For tumor growth: use female rats with N-nitrosomethylurea-induced mammary tumors. (2) Administer cis-4-hydroxy-L-proline IP (50-200 mg/kg) daily for 14-28 days in sterile saline/PBS. (3) Monitor tumor volume twice weekly. (4) At endpoint, excise tumors, weigh, process for histology (H&E, collagen staining). (5) For collagen metabolism: use 6-8 week old male rats or mice. (6) Administer compound IP (100-200 mg/kg) daily for 3-7 days. (7) Collect skin, tendon, lung, liver, determine collagen content by hydroxyproline assay (hydrolyze in 6M HCl, 110degC 18-24 h, neutralize, react with Chloramine T/Ehrlich's reagent, OD560). (8) For fibrosis models (bleomycin, CCl4), administer compound (50-200 mg/kg IP daily) during disease induction, measure collagen deposition by histology (Sirius Red) and hydroxyproline.
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| ADME/Pharmacokinetics |
Soluble in water (≥50 mg/mL) and 0.1M HCl. For cell culture, dissolve directly or prepare 100-500 mM stock in water, filter sterilize. For in vivo, dissolve in sterile saline or PBS at 20-100 mg/mL. Storage: powder at -20degC for 3 years; solution at -20degC for 6 months, -80degC for 1 year. PK in rodents (IP 100 mg/kg): plasma Cmax ~1-2 mM, t1/2 ~1-2 h, rapidly excreted in urine. Oral bioavailability ~20-40%.
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| Toxicity/Toxicokinetics |
In vitro: CCK-8 on fibroblasts or HEK293 cells, IC50 ~10-20 mM. In vivo: acute IP LD50 in mice ~500-1000 mg/kg. High doses (200-500 mg/kg IP) cause lethargy, weight loss, GI distress. Chronic (50-200 mg/kg/day for 2-4 weeks) tolerated but may decrease collagen synthesis and impair wound healing. No significant hepatotoxicity or nephrotoxicity. For research use only.
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| References |
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| Additional Infomation |
cis-4-hydroxy-L-proline is a derivative of L-proline in which the hydrogen at the 4-position of the pyrrolidine ring is replaced by a hydroxyl group (S-configuration). It is a metabolite, belonging to the non-protein L-α-amino acid family, and is also a 4-hydroxyproline. It is a zwitterionic tautomer of cis-4-hydroxy-L-proline. Cis-4-hydroxyproline has been reported to exist in fescue (Festuca ovina), meadow fescue (Festuca pratensis), and other organisms with relevant data. It is the hydroxylated form of the imino acid proline. Ascorbic acid deficiency leads to impaired hydroxyproline production.
cis-4-Hydroxy-L-proline is a proline analog incorporated into procollagen instead of proline, producing non-helical procollagen that fails to fold properly, leading to ER retention and increased intracellular degradation. It is a potent inhibitor of collagen secretion and deposition. Used as a research tool to study collagen biosynthesis, folding, and fibrotic diseases (pulmonary fibrosis, liver cirrhosis, scleroderma). Also shows anti-tumor activity in rat mammary tumor model. Not FDA-approved; strictly for research use. |
| Molecular Formula |
C5H9NO3
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|---|---|
| Molecular Weight |
131.13
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| Exact Mass |
131.058
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| CAS # |
618-27-9
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| Related CAS # |
cis-4-Hydroxy-L-proline-d3
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| PubChem CID |
440015
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
355.2±42.0 °C at 760 mmHg
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| Melting Point |
257ºC
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| Flash Point |
168.6±27.9 °C
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| Vapour Pressure |
0.0±1.8 mmHg at 25°C
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| Index of Refraction |
1.540
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| LogP |
-1.84
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| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
9
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| Complexity |
125
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C1[C@@H](CN[C@@H]1C(=O)O)O
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| InChi Key |
PMMYEEVYMWASQN-IMJSIDKUSA-N
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| InChi Code |
InChI=1S/C5H9NO3/c7-3-1-4(5(8)9)6-2-3/h3-4,6-7H,1-2H2,(H,8,9)/t3-,4-/m0/s1
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| Chemical Name |
(2S,4S)-4-hydroxypyrrolidine-2-carboxylic acid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O :~100 mg/mL (~762.60 mM)
DMSO :< 1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (762.60 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 7.6260 mL | 38.1301 mL | 76.2602 mL | |
| 5 mM | 1.5252 mL | 7.6260 mL | 15.2520 mL | |
| 10 mM | 0.7626 mL | 3.8130 mL | 7.6260 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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