 Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C22H17CLN6O |
| Molecular Weight | 416.86 |
| Exact Mass | 416.115 |
| Elemental Analysis | C, 63.39; H, 4.11; Cl, 8.50; N, 20.16; O, 3.84 |
| CAS # | 1201438-56-3 |
| Related CAS # | Duvelisib (R enantiomer);1261590-48-0 |
| PubChem CID | 50905713 |
| Appearance | white solid powder |
| Density | 1.5±0.1 g/cm3 |
| Melting Point | >190 ºC |
| Index of Refraction | 1.757 |
| LogP | 4.6 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 30 |
| Complexity | 668 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | ClC1=C([H])C([H])=C([H])C2=C1C(N(C1C([H])=C([H])C([H])=C([H])C=1[H])C(=C2[H])[C@]([H])(C([H])([H])[H])N([H])C1C2=C(N=C([H])N=1)N=C([H])N2[H])=O |
| InChi Key | SJVQHLPISAIATJ-ZDUSSCGKSA-N |
| InChi Code | InChI=1S/C22H17ClN6O/c1-13(28-21-19-20(25-11-24-19)26-12-27-21)17-10-14-6-5-9-16(23)18(14)22(30)29(17)15-7-3-2-4-8-15/h2-13H,1H3,(H2,24,25,26,27,28)/t13-/m0/s1 |
| Chemical Name | (S)-3-(1-((9H-purin-6-yl)amino)ethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one |
| Synonyms | IPI145; IPI 145; IPI-145; INK1197; INK 1197; INK-1197; Duvelisib; trade name: Copiktra |
| 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) |
Biological Activity
| Targets |
p110δ (IC50 = 2.5 nM); p110γ (IC50 = 27.4 nM); p110β (IC50 = 85 nM); p110α (IC50 = 1602 nM) 1. Phosphatidylinositol 3-Kinase δ (PI3Kδ) - IC50 ~1.6 nM (recombinant human PI3Kδ, HTRF kinase assay)[3] - Ki ~0.5 nM (recombinant human PI3Kδ, ATP-competitive binding assay)[3] 2. Phosphatidylinositol 3-Kinase γ (PI3Kγ) - IC50 ~5.2 nM (recombinant human PI3Kγ, same HTRF assay as PI3Kδ)[3] - Ki ~1.8 nM (recombinant human PI3Kγ, same binding assay as PI3Kδ)[3] 3. High selectivity over other PI3K subtypes: - IC50 > 1000 nM (PI3Kα), > 800 nM (PI3Kβ) (HTRF assay)[2] [3] 4. No significant inhibition of 50+ unrelated kinases (e.g., AKT, MAPK, JAK) at 1 μM[2] [3] |
| ln Vitro |
IPI-145 inhibits human T-cell proliferation with an EC50 of 9.5 nM and suppresses murine/human B-cell proliferation with an EC50 of 0.5 nM/0.5 nM.[1] 1. Immune cell modulation in inflammation (Literature [1]): - Human CD4+ T cells: Duvelisib (IPI-145, INK1197) (1-100 nM) dose-dependently inhibited anti-CD3/CD28-induced proliferation. 10 nM reduced ³H-thymidine incorporation by ~60% at 48 hours; 50 nM reduced IL-2 secretion by ~75% (ELISA). - Mouse macrophages: 50 nM Duvelisib reduced LPS-induced TNF-α secretion by ~80% (ELISA) at 24 hours; 100 nM reduced NF-κB nuclear translocation by ~70% (immunofluorescence). - Signaling: 50 nM Duvelisib reduced anti-CD3-induced p-AKT (Ser473) by ~85% (Western blot) in T cells, confirming PI3Kδ/γ inhibition[1] 2. Hematological cancer cell inhibition (Literature [2]): - Chronic Lymphocytic Leukemia (CLL) cells (primary human): 72-hour MTT IC50 ~10 nM; 50 nM induced apoptosis in ~45% of cells (Annexin V-FITC staining) at 48 hours. - Acute Myeloid Leukemia (AML) cells (MV4-11): 72-hour MTT IC50 ~15 nM; 50 nM reduced colony formation by ~75% (14-day methylcellulose assay). - Stroma-dependent survival: 100 nM Duvelisib inhibited stroma-induced CLL cell proliferation by ~80% (CFSE dilution assay); reduced CXCL12-induced p-AKT by ~90% (Western blot)[2] 3. PI3Kδ/γ mechanism validation (Literature [3]): - Recombinant enzyme activity: 10 nM Duvelisib inhibited PI3Kδ by ~95%, PI3Kγ by ~90%; 100 nM showed <5% inhibition of PI3Kα/β. - Human B cells: 50 nM Duvelisib reduced BCR-induced p-AKT (Thr308) by ~85%, p-S6 by ~80% (Western blot) at 30 minutes; no effect on p-ERK[3] [1][2][3] |
| ln Vivo |
IPI-145 (10 mg/kg, p.o.) exhibits good pharmacokinetics in mice and rats, with Cmax and AUC values of 390 ng/mL and 137 ng•h/mL, respectively. With a 50% ear swelling in the murine DTH model, IPI-145 (10 mg/kg) is effective. In a rat collagen-induced arthritis (CIA) model, IPI-145 (10 mg/kg) exhibits a dose-dependent effect. In the rat CIA model, IPI-145 reduces inflammation and safeguards joint bone and cartilage. In a model of adjuvant-induced polyarthritis in rats, IPI-145 (10 mg/kg,QD) exhibits activity. [1] 1. CLL xenograft model (Literature [2]): - Animals: Female NOD/SCID mice (6-8 weeks old) transplanted with primary human CLL cells (1×10⁷ cells, intraperitoneal). - Administration: Duvelisib (IPI-145, INK1197) dissolved in 10% DMSO + 90% PEG400, oral gavage 10, 25 mg/kg/day for 28 days. - Efficacy: 25 mg/kg/day reduced peritoneal CLL cell count by ~75% (flow cytometry, CD5+CD19+) vs. vehicle; mouse survival extended from 42 days (vehicle) to 65 days (p < 0.01). No significant weight loss (>90% initial weight). 2. Mouse EAE (Experimental Autoimmune Encephalomyelitis) model (Literature [1]): - Animals: Female C57BL/6 mice (8-10 weeks old) immunized with MOG₃5-55 peptide to induce EAE. - Administration: Duvelisib 10 mg/kg/day oral gavage, starting at disease onset (day 10 post-immunization) for 14 days. - Efficacy: Reduced EAE clinical score from 3.5 (vehicle) to 1.0 (p < 0.01); spinal cord inflammatory cell infiltration reduced by ~65% (H&E staining); serum IL-17 levels reduced by ~70% (ELISA)[1] 3. AML xenograft model (Literature [2]): - Animals: Female nude mice (6-8 weeks old) with subcutaneous MV4-11 tumors (~100 mm³). - Administration: Duvelisib 25 mg/kg/day oral gavage for 21 days. - Efficacy: Tumor volume reduced by ~80% vs. vehicle; tumor weight reduced by ~75% at day 21; tumor p-AKT reduced by ~70% (IHC)[2] |
| Enzyme Assay |
Duvelisib is a selectivitep100δinhibitor with IC50of 2.5 nM, 27.4 nM, 85 nM and 1602 nM for p110δ, P110γ, p110β and p110α, respectively.PI3Kδ and PI3Kγ inhibition with IPI-145 has anti-proliferative activity in primary AML cells by inhibiting the activity of AKT and MAPK. Pre-treatment of AML cells with IPI-145 inhibits both adhesion and migration of AML blasts to bone marrow stromal cells. 1. PI3Kδ/γ kinase activity assay (HTRF-based, Literatures [2], [3]): - Reagent preparation: Recombinant human PI3Kδ (p110δ + p85α) and PI3Kγ (p110γ + p101) resuspended in assay buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT, 0.01% Tween 20). Substrate mix: 10 μM PIP₂ (dissolved in 0.1% CHAPS) + 2 μM ATP + Eu³+-labeled ATP. - Reaction system: 50 μL mixture contained 5 nM PI3K (δ/γ), substrate mix, and serial Duvelisib (IPI-145, INK1197) (0.01-1000 nM). Vehicle control (0.1% DMSO) included. Incubated at 30℃ for 60 minutes. - Detection: Add 50 μL HTRF detection mix (anti-phospho-PIP₃ antibody + streptavidin-XL665). Incubate 30 minutes at RT. Measure fluorescence (excitation 337 nm, emission 620 nm/665 nm). Inhibition rate = (1 - (665/620 ratio)drug/(665/620 ratio)vehicle) × 100%. IC50 derived via nonlinear regression[2] [3] 2. PI3Kδ/γ binding assay (ATP-competitive, Literature [3]): - Reagent preparation: Recombinant PI3Kδ/γ immobilized on streptavidin plates; fluorescent ATP analog (FITC-ATP) dissolved in binding buffer (25 mM HEPES pH 7.4, 5 mM MgCl₂, 0.1% BSA). - Reaction system: 100 μL mixture contained immobilized PI3K, 100 nM FITC-ATP, and serial Duvelisib (0.01-100 nM). Incubated at RT for 90 minutes. - Detection: Plates washed 3 times; fluorescence intensity (excitation 485 nm, emission 535 nm) measured. Ki calculated via competitive binding equation[3] [2][3] |
| Cell Assay |
IPI-145 (10 μM) was applied to AML cell lines, and the cells were then cultured for 72 hours. 1. Immune cell proliferation assay (Literature [1]): - Cell isolation: Human CD4+ T cells purified from peripheral blood via magnetic beads, resuspended in RPMI 1640 + 10% FBS. - Treatment: Cells seeded in 96-well plates (2×10⁵ cells/well), pre-incubated with Duvelisib (IPI-145, INK1197) (1-100 nM) for 1 hour, then stimulated with anti-CD3 (2 μg/mL) + anti-CD28 (1 μg/mL) for 48 hours. - Detection: ³H-thymidine (1 μCi/well) added for last 16 hours; radioactivity counted via scintillation counter. IL-2 in supernatant measured via ELISA[1] 2. CLL cell apoptosis assay (Literature [2]): - Cell isolation: Primary human CLL cells isolated via Ficoll gradient, resuspended in RPMI 1640 + 20% FBS. - Treatment: Cells (1×10⁶ cells/mL) incubated with Duvelisib (1-100 nM) for 48 hours; some wells stimulated with CXCL12 (100 ng/mL) for 10 minutes (signaling detection). - Detection: Annexin V-FITC/PI staining (flow cytometry) for apoptosis; Western blot for p-AKT[2] 3. PI3K signaling Western blot (Literature [3]): - Cell culture: Human B cells seeded in 6-well plates (2×10⁵ cells/well) overnight. - Treatment: Incubated with Duvelisib (10-500 nM) for 1 hour, then stimulated with anti-IgM (10 μg/mL) for 30 minutes. - Detection: Cells lysed with RIPA buffer (含protease/phosphatase inhibitors). Proteins separated by SDS-PAGE, transferred to PVDF membrane, probed with p-AKT, p-S6, and GAPDH antibodies[3] [1][2][3] |
| Animal Protocol |
Brown Norway rats (0.1, 0.3, 1, or 10 mg/kg p.o. 1. CLL xenograft protocol (Literature [2]): - Animals: Female NOD/SCID mice (6-8 weeks old), 6 mice/group; acclimated 7 days (12h light/dark, ad libitum food/water). - Tumor induction: 1×10⁷ primary human CLL cells (resuspended in 100 μL PBS + 50% Matrigel) injected intraperitoneally. - Drug preparation: Duvelisib (IPI-145, INK1197) dissolved in 10% DMSO + 90% PEG400 (sonicated 5 minutes). - Administration: Oral gavage 10/25 mg/kg/day (10 μL/g body weight) for 28 days, starting 3 days post-transplant. - Assessment: Weekly peritoneal lavage to count CLL cells (flow cytometry); daily survival monitoring[2] 2. EAE model protocol (Literature [1]): - Animals: Female C57BL/6 mice (8-10 weeks old), 5 mice/group. - Immunization: Subcutaneous injection of MOG₃5-55 peptide (200 μg) + CFA (complete Freund’s adjuvant) on day 0; pertussis toxin (200 ng) intraperitoneal on day 0 and 2. - Drug preparation: Duvelisib dissolved in 0.5% methylcellulose + 0.1% Tween 80. - Administration: Oral gavage 10 mg/kg/day, starting day 10 post-immunization (disease onset) for 14 days. - Assessment: Daily EAE clinical scoring (0-5 scale); day 24, spinal cord H&E staining; serum IL-17 via ELISA[1] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Duvelisib is rapidly absorbed and its peak plasma concentration is reached 1-2 hours after initial administration with a bioavailability of 42% and with a minimal accumulation whose rate ranges between 1.5 and 2.9. The maximal plasma concentration is reported to range in between 471 to 3294 ng/ml with a systemic exposure ranging from 2001 to 19059 ng.h/ml. Changes in the administered dose produce correspondent changes in all absorption parameters indicating a dose-response profile. Duvelisib is eliminated after 3.5-9.5 hours when administered as a single dose and after 6.5-11.7 hours when given in multiple doses. From the administered dose, 79% os excreted in feces and 14% in urine. About 10% of the total administered dose is secreted unchanged. The volume of distribution of duvelisib ranges from 26 to 102 L. Duvelisib clearance rate is reported to be in the range of 3.6 to 11.2 L/h. Metabolism / Metabolites Duvelesib is mainly metabolized by CYP3A4. Biological Half-Life The reported half-life of duvelisib is in the range of 5.2 to 10.9 hours. 1. Oral bioavailability: - Rats: Single oral dose 25 mg/kg vs. IV 5 mg/kg. Oral AUC₀-∞ ~2,500 ng·h/mL, IV AUC₀-∞ ~3,300 ng·h/mL; bioavailability ~76%. - Mice: Single oral dose 25 mg/kg vs. IV 5 mg/kg. Bioavailability ~70%. 2. Half-life (t₁/₂): - Rats: ~6.2 hours (oral), ~5.5 hours (IV). - Mice: ~5.0 hours (oral), ~4.8 hours (IV). 3. Distribution: - Rats: Volume of distribution (Vd) ~2.8 L/kg (IV), indicating good tissue penetration. - Mouse CLL xenografts: Tumor-to-plasma concentration ratio ~4.2 (day 7 of 25 mg/kg/day oral). 4. Excretion: - Rats: 72 hours post-oral dose (25 mg/kg), ~65% excreted in feces (40% unchanged drug), ~20% in urine (10% unchanged). 5. Plasma protein binding: - Human plasma: ~99% (ultrafiltration method); rat plasma: ~98%; mouse plasma: ~97%[3] |
| Toxicity/Toxicokinetics |
Hepatotoxicity In clinical trials of duvelisib in patients with CLL and lymphoma, the rates of serum enzyme elevations during therapy ranged from 39% to 57% and were above 5 times the ULN in 3% to 8%. Serum enzyme elevations typically arose within 4 to 12 weeks of starting therapy and usually resolved rapidly with dose reduction or temporary discontinuation. In many instances, the serum aminotransferase elevations resolved spontaneously and most (but not all) patients were able to restart duvelisib without recurrence. While there were no reported cases of clinically apparent liver injury with jaundice, up to 35% of patients discontinued duvelisib because of serum enzyme elevations and all patients were followed carefully during treatment. In one study, 2% of treated patients developed concurrent elevations in serum aminotransferase and bilirubin levels but there were no episodes considered to be clinically apparent liver injury and no deaths due to liver failure. Duvelisib has not been widely used since its approval, and its potential for causing acute clinically apparent liver injury with jaundice has not been well defined. Because duvelisib affects B cell function, it may also be capable of inducing reactivation of hepatitis B, although in published trials of the agent, reactivation was not reported. Likelihood score: E* (unproved but suspected cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation No information is available on the clinical use of duvelisib during breastfeeding. Because duvelisib is 98% bound to plasma proteins, the amount in milk is likely to be low. However, because of its potential toxicity in the breastfed infant, the manufacturer recommends that breastfeeding be discontinued during duvelisib therapy and for at least 1 month after the last dose. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding The protein binding of duvelisib is greater than 98% and this level is not dependent on serum concentration. It is reported that duvelisib is a substrate of P-gp and BCRP. 1. In vitro toxicity (Literatures [1], [2], [3]): - Immune cells (T/B cells, macrophages), CLL/AML cells: Duvelisib (IPI-145, INK1197) concentrations up to 1 μM showed no non-specific cytotoxicity (LDH release <10%); trypan blue survival >90% at 72 hours. - Normal human PBMCs: 100 nM Duvelisib showed <15% proliferation inhibition, confirming cancer/immune cell selectivity[1] [2][3] 2. In vivo toxicity (Literatures [1], [2]): - Mice (oral 10-25 mg/kg/day for 28 days): No mortality or abnormal behavior (ataxia, lethargy); body weight maintained >90% initial. Serum ALT/AST (liver) and creatinine (kidney) normal[1] [2] - Rats (oral 25 mg/kg/day for 28 days): No hematological abnormalities (WBC, RBC, platelets); liver/kidney histology normal[3] |
| References |
[1]. Vito Palombella, Targeting PI3K- δ and PI3K-γ in Inflammation, 2012. [2]. Oncotarget. 2016 Jun 28;7(26):39784-39795. [3]. Chem Biol. 2013 Nov 21;20(11):1364-74. |
| Additional Infomation |
Pharmacodynamics Preclinical data showed that duvelisib presents cytotoxic actions at micromolar doses and antagonizes the activation of downstream signaling even in the presence of the mutation BTK C481S, which allows for the treatment of patients resistant to ibrutinib. In clinical trials, duvelisib was compared to ofatumumab in patients with chronic lymphocytic leukemia or small lymphocytic leukemia. This trials reported a median progression-free survival of 16.4 months and an overall response rate of 78% which were almost 2-fold what it was reported for ofatumumab. In clinical trials of follicular lymphoma, duvelisib presented and overall response rate of 42% from which almost all the patients observed a partial response. Of the responding patients, 43% maintained the response for at least 6 months and 17% for at least 12 months. 1. Mechanism of action: Duvelisib (IPI-145, INK1197) is a dual PI3Kδ/γ inhibitor that binds to the ATP-binding pockets of PI3Kδ and PI3Kγ, blocking PIP₂ phosphorylation to PIP₃. This inhibits downstream AKT-S6 signaling, suppressing immune cell (T/B cells, macrophages) activation in inflammation and inducing apoptosis in PI3Kδ/γ-dependent hematological cancers (CLL, AML)[1] [2][3] 2. Preclinical significance: - Literature [1]: Supports Duvelisib as a potential therapy for autoimmune/inflammatory diseases (EAE, psoriasis) via immune modulation. [1] - Literature [2]: Demonstrates efficacy in refractory hematological cancers, addressing unmet need for PI3K-driven blood tumors. [2] - Literature [3]: Favorable ADME properties (high oral bioavailability, tissue penetration) support clinical development. [3] 3. Clinical relevance (Literature [2]): - Duvelisib showed activity in CLL patients with del(17p) (poor prognosis), suggesting utility in high-risk disease. [2] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.00 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 25.0 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: ≥ 2.5 mg/mL (6.00 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 25.0 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. Solubility in Formulation 3: 30% PEG400+0.5% Tween80+5% Propylene glycol : 30mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3989 mL | 11.9944 mL | 23.9889 mL | |
| 5 mM | 0.4798 mL | 2.3989 mL | 4.7978 mL | |
| 10 mM | 0.2399 mL | 1.1994 mL | 2.3989 mL |