| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| 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 |
- 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]. Zetomipzomib maleate is an immunoproteasome selective inhibitor [3]. Additionally, constitutive proteasome β5 subunit (IC50=688 nM) and MECL-1 subunit (IC50=623 nM) are inhibited by methotrimib maleate. LMP7 and LMP2 in MOLT-4 cells are selectively inhibited by methotrimib maleate. Peripheral blood mononuclear cells (PBMCs) and methotrimib maleate (250 nM) exhibit a similar cytokine inhibitory profile [1]. |
| ln Vivo |
- 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]. In an anti-collagen antibody-induced arthritis (CAIA) animal, methotrimib maleate (5 mg/kg; intravenously; repeated doses on days 6, 8, 11, and 13) has demonstrated effectiveness[1]. |
| 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 |
- 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]. Animal/Disease Models: 7-8 week old female balb/c (Bagg ALBino) mouse (CAIA model)[1] Doses: Iv; Dosing was repeated on days 6, 8, 11, and 13 until for 15 day Route of Administration: 5 mg/kg Experimental Results: demonstrated efficacy in the anticollagen antibody induced arthritis (CAIA) model. |
| 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 |
|
| Additional Infomation |
- 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 |
C34H46N4O12
|
|---|---|
| Molecular Weight |
702.75
|
| Exact Mass |
702.311
|
| Elemental Analysis |
C, 58.11; H, 6.60; N, 7.97; O, 27.32
|
| CAS # |
2170983-62-5
|
| Related CAS # |
Zetomipzomib;1629677-75-3
|
| PubChem CID |
162640583
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
13
|
| Rotatable Bond Count |
16
|
| Heavy Atom Count |
50
|
| Complexity |
1130
|
| Defined Atom Stereocenter Count |
5
|
| SMILES |
C(/C(=O)O)=C/C(=O)O.C([C@@]1(OC1)C)(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)CN1CCOCC1)[C@@H](C1C=CC(OC)=CC=1)O)CC1=CCCC1
|
| InChi Key |
HVKAUVYFVFFXLW-MTOCQVLVSA-N
|
| InChi Code |
InChI=1S/C30H42N4O8.C4H4O4/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;5-3(6)1-2-4(7)8/h6,8-11,19,23,25-26,36H,4-5,7,12-18H2,1-3H3,(H,31,35)(H,32,39)(H,33,38);1-2H,(H,5,6)(H,7,8)/b;2-1-/t19-,23-,25-,26+,30+;/m0./s1
|
| Chemical Name |
(Z)-but-2-enedioic acid;(2S,3R)-N-[(2S)-3-(cyclopenten-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-hydroxy-3-(4-methoxyphenyl)-2-[[(2S)-2-[(2-morpholin-4-ylacetyl)amino]propanoyl]amino]propanamide
|
| Synonyms |
KZR-616 maleate; Zetomipzomib Maleate; KZR616 maleate; 2170983-62-5; KZR 616 maleate; Zetomipzomib (maleate); Zetomipzomib maleate (USAN)
|
| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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: 250 mg/mL (355.75 mM)
H2O: 50 mg/mL (71.15 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (2.96 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (2.96 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: 100 mg/mL (142.30 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.4230 mL | 7.1149 mL | 14.2298 mL | |
| 5 mM | 0.2846 mL | 1.4230 mL | 2.8460 mL | |
| 10 mM | 0.1423 mL | 0.7115 mL | 1.4230 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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