| Size | Price | Stock | Qty |
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| 500mg |
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| 1g | |||
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Zetomipzomib (KZR-616) is a potent, peptide epoxyketone-based, and selective dual inhibitor of immunoproteasome subunit LMP7 (low molecular mass polypeptide-7)/LMP2 (multicatalytic endopeptidase complex subunit-1) with IC50 of 39 nM/139 nM . Selective immunoproteasome inhibition is a promising approach for treating autoimmune disorders. KZR-616 has anti-Inflammatory efficacy and is currently in clinical trials for treatment of rheumatic arthritis.
Zetomipzomib (KZR-616) is a novel, selective, and covalent tripeptide epoxyketone inhibitor of the immunoproteasome [1][3].| Targets |
Immunoproteasome subunits LMP7 (β5i) and LMP2 (β1i) [1][2][3].
- IC50 for human LMP7 (β5i): 39 nM [1][3]. - IC50 for human LMP2 (β1i): 623 nM [1][3]. - IC50 for human MECL-1 (β2i): 131 nM [1][3]. - Selectivity ratio (β5c/β5i): 17.6 [1][3]. |
|---|---|
| ln Vitro |
Additionally, constitutive proteasome β5 subunit (IC50=688 nM) and MECL-1 subunit (IC50=623 nM) are inhibited by zestomipzomib. In MOLT-4 cells, zetomipzomib continues to selectively inhibit LMP7 and LMP2. Similar cytokine inhibitory effects are seen by peripheral blood mononuclear cells (PBMC) and methotrimipzomib (250 nM) [1]. Based on ONX-0914 and PR-924, zetomipzomib is an immunoproteasome selective inhibitor that has been optimized and determined [3].
- Cytokine Inhibition in Human PBMCs and T cells: At a concentration resulting in inhibition of LMP7 by 89% and LMP2 by 59%, KZR-616 induced a decrease in pro-inflammatory cytokine production in human peripheral blood mononuclear cells (PBMCs) stimulated with endotoxin [2]. It blocked T-cell production of IFN-γ, TNF-α, and GM-CSF from CD4+ T cells stimulated with antibodies to CD3 and CD28 [2]. - Inhibition of B-cell Differentiation: KZR-616 inhibited the differentiation of human B-cells into plasmablasts when stimulated with IL-21 and antibodies to CD40 and IgM [2]. - Protease Selectivity: As expected of compounds from the epoxyketone chemical class, KZR-616 showed no inhibition at 10 µM against a broad selectivity panel of 20 serine, metallo-, cysteine, and aspartyl proteases and 11 hydrolases [1]. |
| ln Vivo |
In a model of anti-collagen antibody-induced arthritis (CAIA), methotrimipzomib (5 mg/kg; intravenously; repeated doses on days 6, 8, 11, and 13) has demonstrated effectiveness [1].
- Efficacy in SLE Mouse Model (NZB/W F1): KZR-616 treatment in diseased mice resulted in a complete resolution of proteinuria and significant reductions in autoantibody production and renal IgG deposition [2]. The halt in disease progression was durable, as proteinuria levels did not significantly increase 8 weeks after treatment discontinuation [2]. Histologic analysis following 12 weeks of treatment revealed a complete prevention in glomerulonephritis and sclerosis [2]. - Combination Therapy: Administration of KZR-616 in combination with mycophenolate mofetil (MMF) resulted in significantly greater disease inhibition and prolonged survival compared to either treatment alone in the NZB/W F1 model [2]. - Immune Cell Depletion: Levels of activated T- and B-cells and short- and long-lived plasma cells were effectively depleted in diseased animals following KZR-616 treatment [2]. - Efficacy in Collagen Antibody Induced Arthritis (CAIA) Model: KZR-616 showed comparable efficacy to ONX 0914 at half the dosage (5 mg/kg vs. 10 mg/kg for ONX 0914) in the CAIA model [1]. Subcutaneous administration gave comparable results at similar dosages [1]. - Effect on T-cell Dependent Antibody Response (TDAR): KZR-616 had no significant effect on TDAR in mice or monkeys and did not affect the number of circulating lymphocytes in monkeys [2]. |
| Enzyme Assay |
- Proteasome Constitutive/Immunoproteasome Subunit ELISA (ProCISE): This ELISA-based technique was utilized for quantitative assessment of subunit-specific activity [1].
For cell lysate analysis, test compounds were serially diluted in DMSO, then diluted in hypotonic lysis buffer [1]. MOLT-4 or A20 cell lysate was treated with the compound for 1 hour at 25°C [1]. Treated cell lysate was incubated with a biotinylated proteasome active site binding probe for 2 hours at 25°C [1]. Subunits bound to the probe were isolated with streptavidin-conjugated sepharose beads [1]. After multiple rinses, the activity was measured [1]. This method was used to determine the IC50 values for KZR-616 against various proteasome subunits [1]. |
| Cell Assay |
- PBMC Whole Cell Assay for Subunit Inhibition and Cytokine Analysis: Cryopreserved PBMCs from healthy volunteers were cultured [1].
Compounds were diluted in DMSO and then in cell growth media [1]. Live PBMCs were treated with compounds for 1 hour at 37°C, then washed [1]. To measure immunoproteasome subunit inhibition, cells were frozen, thawed, and processed to lysate, and the ProCISE assay was performed as described in the Enzyme Assay section [1]. To measure cytokine inhibition, PBMCs were stimulated with LPS (1 µg/mL) or antibodies against CD3 and CD28 for 24 hours after compound treatment [1]. Supernatants were analyzed for TNFα, IL-6, the p40 subunit of IL-12/23, and IFNγ via electrochemiluminescent ELISA [1]. - B-cell Plasmablast Differentiation Assay: Human B-cells were stimulated with IL-21 and antibodies to CD40 and IgM [2]. KZR-616 was added, and its effect on the differentiation of B-cells into plasmablasts was measured [2]. |
| Animal Protocol |
Animal/Disease Models: 7-8 weeks old female balb/c (Bagg ALBino) mouse (CAIA model) [1]
Doses: intravenous (iv) (iv)injection; repeated dosing on days 6, 8, 11 and 13 until day 15 Dosing: 5mg/kg Experimental Results: Demonstrated efficacy in anti-collagen antibody-induced arthritis (CAIA) model. - Mouse Pharmacodynamic Study: BALB/c mice were administered KZR-616 intravenously (iv) at 5 mg/kg [1]. Kidney and splenocyte (erythrocyte-depleted) samples were taken 1 hour after dosing [1]. The activity of LMP7, LMP2, MECL-1 (in splenocytes), and β5 (in kidney) was measured by ProCISE [1]. Data were normalized to the average activity of vehicle-treated animals [1]. - Mouse Collagen Antibody Induced Arthritis (CAIA) Model: BALB/c mice received 1.75 mg of a cocktail of five antibodies against type II collagen on day 0, followed by 25 µg of LPS on day 3 [1]. On day 4, when disease symptoms were present, animals were randomized and treated iv with vehicle, KZR-616 (at 2.5 or 5 mg/kg), or ONX 0914 (10 mg/kg) [1]. Dosing was repeated on days 6, 8, 11, and 13 [1]. Clinical scores (0-4 per paw; n=7/group) were followed until day 15 [1]. - Mouse SLE Model (NZB/W F1): The therapeutic effect of KZR-616 alone or in combination with mycophenolate mofetil (MMF) was evaluated in the NZB/W F1 model of SLE [2]. Dosing regimens and administration routes (iv or subcutaneous) were described as effective at well-tolerated doses, enabling durable disease remission [2]. |
| ADME/Pharmacokinetics |
- General Pharmacokinetic Profile: Peptide epoxyketones, including KZR-616, show rapid pharmacokinetic clearance across species, with a typical half-life (t1/2) of less than 15 minutes [1].
Like other covalent inhibitors, their pharmacodynamic effects outlast compound pharmacokinetics and reflect the turnover of the covalently inhibited enzyme in perfused tissues [1]. - Mouse PK: The pharmacokinetics of KZR-616 upon intravenous administration to BALB/c mice were comparably rapid (t1/2 < 7 min, clearance > 73 mL min-1 kg-1) [1]. |
| Toxicity/Toxicokinetics |
- Tolerability: Based on body weights and cage-side observations, no change in tolerability was detected between any treatment group (vehicle vs. KZR-616) in the CAIA mouse model [1].
- Lack of Immunosuppression: KZR-616 had no significant effect on T-cell dependent antibody responses (TDAR) in mice or monkeys and did not affect the number of circulating lymphocytes in monkeys, suggesting it does not cause general immunosuppression [2]. - Cell Viability: At fully inhibitory concentrations of all three immunoproteasome subunits, greater cytokine inhibition was achieved but was also associated with a loss of viability in the cells [1]. |
| References |
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| Additional Infomation |
Zetomipzomib is being investigated in the clinical trial NCT04033926 (a phase 2 study evaluating the safety and efficacy of KZR-616 in patients with active polymyositis or dermatomyositis).
- Clinical Status: KZR-616 is the first immunoproteasome-selective inhibitor to enter clinical trials [1][3]. It is being developed for the treatment of autoimmune diseases, including systemic lupus erythematosus (SLE) and lupus nephritis [1][2][3]. A Phase 1b/2 study was listed for patients with SLE (NCT03393013) [1][3]. - Mechanism of Action: Studies demonstrated that a required immunoproteasome subunit inhibition profile for anti-inflammatory efficacy is the dual inhibition of LMP7/LMP2 or LMP7/MECL-1 [1]. Selective LMP7 inhibition alone (by compound 8) was insufficient to potently inhibit cytokine expression or disease progression [1]. KZR-616 was designed to achieve this dual inhibition profile [1]. - Structural Optimization: KZR-616 was optimized from earlier leads (ONX 0914, PR-924) by incorporating a P1 cyclopentene ring and an R-hydroxyl group substitution at the β position of the P2 methyltyrosine side chain [1]. This resulted in vastly improved solubility (2.5-fold over the analogue 11) while maintaining the desired subunit inhibition profile and efficacy [1]. |
| Molecular Formula |
C30H42N4O8
|
|---|---|
| Molecular Weight |
586.676488399506
|
| Exact Mass |
586.3
|
| CAS # |
1629677-75-3
|
| Related CAS # |
Zetomipzomib maleate;2170983-62-5
|
| PubChem CID |
117607904
|
| Appearance |
White to off-white solid at room temperature
|
| LogP |
0.4
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
14
|
| Heavy Atom Count |
42
|
| Complexity |
1010
|
| Defined Atom Stereocenter Count |
5
|
| SMILES |
O1C[C@]1(C)C([C@H](CC1=CCCC1)NC([C@H]([C@@H](C1C=CC(=CC=1)OC)O)NC([C@H](C)NC(CN1CCOCC1)=O)=O)=O)=O
|
| InChi Key |
GHYOCDFICYLMRF-UTIIJYGPSA-N
|
| InChi Code |
InChI=1S/C30H42N4O8/c1-19(31-24(35)17-34-12-14-41-15-13-34)28(38)33-25(26(36)21-8-10-22(40-3)11-9-21)29(39)32-23(16-20-6-4-5-7-20)27(37)30(2)18-42-30/h6,8-11,19,23,25-26,36H,4-5,7,12-18H2,1-3H3,(H,31,35)(H,32,39)(H,33,38)/t19-,23-,25-,26+,30+/m0/s1
|
| Chemical Name |
(2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-(2-morpholinoacetamido)propanamido)propanamide
|
| Synonyms |
KZR616; KZR-616; Zetomipzomib; 1629677-75-3; O4BT6C02M2; 4,5-Anhydro-1-(1-cyclopenten-1-yl)-1,2-dideoxy-4-C-methyl-2-((N-(2-(4-morpholinyl)acetyl)-L-alanyl-(betaR)-beta-hydroxy-O-methyl-L-tyrosyl)amino)-D-erythro-3-pentulose; KZR 616
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO:~50 mg/mL (85.2 mM)
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7045 mL | 8.5225 mL | 17.0451 mL | |
| 5 mM | 0.3409 mL | 1.7045 mL | 3.4090 mL | |
| 10 mM | 0.1705 mL | 0.8523 mL | 1.7045 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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