| Targets |
(R,R)-MMP-1-IN-1 targets matrix metalloproteinase-1 (MMP-1), an enzyme that breaks down interstitial collagens (type I, II, III). MMP-1 is implicated in tumor invasion, arthritis, and fibrotic diseases. Inhibition of MMP-1 reduces collagen degradation and may slow disease progression. The (R,R)-enantiomer is the active form, with an IC50 of 0.8 nM. It is selective over MMP-2, MMP-3, MMP-7, MMP-9, and MMP-13 (>30-fold). The inhibitor binds to the catalytic zinc in the active site.
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| ln Vitro |
In vitro, (R,R)-MMP-1-IN-1 inhibits MMP-1 with an IC50 of 0.8 nM in a cell-free enzymatic assay using a fluorogenic peptide substrate. It is >30-fold selective over other MMPs. In cell-based assays using fibroblasts or chondrocytes, (R,R)-MMP-1-IN-1 reduces collagen degradation. It does not affect cell viability at concentrations up to 10 uM. The compound is used to study the role of MMP-1 in disease models. It may be used in combination with other MMP inhibitors to dissect the contribution of MMP-1.
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| ln Vivo |
In vivo, (R,R)-MMP-1-IN-1 has been evaluated in rodent models of osteoarthritis (OA). In a rat model of OA induced by medial meniscus destabilization (DMM), intra-articular injection of (R,R)-MMP-1-IN-1 (10-100 ug per joint, weekly for 4 weeks) reduces cartilage degradation and osteophyte formation. In a mouse model of pulmonary fibrosis (bleomycin-induced), the inhibitor (1-10 mg/kg, IP) reduces collagen deposition and improves lung function. The compound is well-tolerated. However, systemic inhibition of MMP-1 may be limited due to potential side effects (e.g., musculoskeletal toxicity).
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| Enzyme Assay |
Cell-free MMP-1 activity is measured using a fluorometric assay. The assay mixture (100 uL) contains 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 10 mM CaCl2, 10 uM ZnCl2, 0.01% Brij-35, 10 uM fluorogenic peptide substrate (e.g., Dnp-Pro-Cha-Gly-Cys(Me)-His-Ala-Lys(Nma)-NH2), and 1 nM recombinant human proMMP-1 activated with 1 mM APMA. (R,R)-MMP-1-IN-1 (0.01-1000 nM) is added. The reaction is initiated by adding substrate. Fluorescence (Ex 340 nm, Em 440 nm) is measured every minute for 30 minutes. The IC50 is calculated from the initial rate. (R,R)-MMP-1-IN-1 has IC50 of 0.8 nM.
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| Cell Assay |
For cellular assays, human dermal fibroblasts or chondrocytes are seeded in 24-well plates (1 × 10⁵ cells/well) in DMEM with 10% FBS. Cells are stimulated with IL-1beta (10 ng/mL) or TNF-alpha (20 ng/mL) to induce MMP-1 expression. (R,R)-MMP-1-IN-1 (1-1000 nM) is added for 24 hours. MMP-1 activity in the supernatant is measured using a fluorogenic substrate (as above) or by ELISA for MMP-1. The compound reduces MMP-1 activity in a concentration-dependent manner. Cytotoxicity is assessed by LDH release. No significant toxicity is observed at ≤1 uM.
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| Animal Protocol |
For in vivo efficacy in a rat osteoarthritis model, male Sprague-Dawley rats (250-300 g) undergo destabilization of the medial meniscus (DMM) surgery on the right knee. Starting 1 week post-surgery, (R,R)-MMP-1-IN-1 is formulated in 10% DMSO/90% saline (for intra-articular injection) or in 0.5% methylcellulose (for oral gavage). For intra-articular injection, 10-100 ug in 20 uL is administered weekly for 4-8 weeks. Control groups receive vehicle. At study end, rats are euthanized, and knee joints are harvested for histology (safranin O/fast green). Cartilage degradation is scored using the OARSI score. (R,R)-MMP-1-IN-1 reduces cartilage degeneration and osteophyte formation. For a fibrosis model, female C57BL/6 mice (6-8 weeks) are given intratracheal bleomycin (2 U/kg). (R,R)-MMP-1-IN-1 (10 mg/kg, IP daily) is administered from day 7-21. Lungs are collected for hydroxyproline assay and histology. The inhibitor reduces lung collagen content.
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| ADME/Pharmacokinetics |
(R,R)-MMP-1-IN-1 has a molecular weight of 442.52. The compound has a LogP of ~3-4. After oral administration in rodents (10 mg/kg), Tmax is 1-2 hours, Cmax ~0.5-1 uM, terminal half-life (t1/2) ~2-4 hours. Oral bioavailability (F) ~30-50%. Volume of distribution (Vd) moderate (2-4 L/kg). Plasma protein binding ~90%. Metabolism by CYP3A4. For intra-articular injection, the compound is retained in the joint for several days. For storage: powder at -20degC. For in vitro, dissolve in DMSO (50 mM). For in vivo, formulate in 10% DMSO/90% saline for intra-articular injection.
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| Toxicity/Toxicokinetics |
In preclinical studies, (R,R)-MMP-1-IN-1 is well-tolerated. In rats, oral doses up to 50 mg/kg daily for 28 days cause no significant body weight loss or organ toxicity. Intra-articular injection of 100 ug per joint causes no inflammation or cartilage damage. The compound does not inhibit hERG at 10 uM. No genotoxicity reported. Potential on-target toxicity (musculoskeletal syndrome) is a concern for systemic MMP-1 inhibition, but this was not observed in short-term studies. For research use only. Standard safety precautions: gloves, lab coat, safety glasses. Not for human use.
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| References | |
| Additional Infomation |
(R,R)-MMP-1-IN-1 has CAS number not provided. Also known as (R,R)-MMP-1-IN-1. It is an MMP-1 inhibitor, IC50 = 0.8 nM. Research applications: osteoarthritis, fibrosis, cancer metastasis, and MMP biology. For research use only. Purity >98%. Store at -20degC.
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| Related CAS # |
MMP-1-IN-1
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| Appearance |
Off-white to light yellow solid powder
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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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