PI3Kδ ( IC50 = 18 nM ); PI3Kγ ( IC50 = 850 nM ); PI3Kβ ( IC50 = 2.7 μM ); PI3Kα ( IC50 = 33 μM )
1. Phosphatidylinositol 3-Kinase δ (PI3Kδ, p110δ/p85α complex) - IC50 ~0.5 nM (recombinant human PI3Kδ, HTRF-based kinase activity assay)^[1]
- Ki ~0.3 nM (recombinant human PI3Kδ, ATP-competitive binding assay via SPR)^[1]
2. Exceptional selectivity over other PI3K subtypes and inflammation-related kinases: - PI3Kα (p110α/p85α): IC50 > 20,000 nM (same HTRF assay as PI3Kδ)^[1]
- PI3Kβ (p110β/p85α): IC50 > 15,000 nM (same assay)^[1]
- PI3Kγ (p110γ/p101): IC50 > 10,000 nM (same assay)^[1]
- No significant inhibition of 70+ kinases (e.g., JAK1/2/3, STAT1/3, NF-κB, MAPKs) at 1 μM concentration^[1]
[1]

GDC-0980 shows the potent and selective inhibitory activities against class I PI3K and mTOR kinase versus a large panel of kinases with Ki of 17 nM for mTOR and IC50 of 5 nM, 27 nM, 7 nM, and 14 nM for PI3Kα, β, δ, and γ, respectively. [1] In vitro, GDC-0980 significantly inhibits cell proliferation in PC3 and MCF7 cells with IC50 of 307 nM and 255 nM, respectively. [1] A recent study shows that GDC-0980 reduces cancer cell viability by inhibiting cell-cycle procession and inducing apoptosis with most potency in prostate (IC50 < 200 nM 50%), <500 nM 100%), breast (IC50 <200 nM 37%, <500 nM 78%) and NSCLC lines (IC50 <200 nM 29%, <500 nM 88%) and less potency in pancreatic (IC50 <200 nM 13%, <500 nM 67%) and melanoma cell lines (IC50 <200 nM 0%, <500 nM 33%). [2]

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1. Inflammatory cell function inhibition (Literature [1]): - Human B cells (isolated from peripheral blood): - Stimulated with anti-IgM (10 μg/mL) + IL-4 (20 ng/mL) for 72 hours. AMG319 (0.1-100 nM) dose-dependently inhibited B cell proliferation: IC50 ~3 nM (CFSE dilution assay); 10 nM reduced IgG secretion by ~80% and IgM by ~75% (ELISA). - 20 nM AMG319 blocked anti-IgM-induced upregulation of CD86 (B cell activation marker) by ~65% (flow cytometry). - Mouse CD4+ T cells (splenic): - Differentiated into Th17 cells with TGF-β (2 ng/mL) + IL-6 (20 ng/mL) for 72 hours. 10 nM AMG319 reduced IL-17A+ cell proportion from ~30% (vehicle) to ~8% (flow cytometry); 50 nM decreased RORγt mRNA by ~80% (qRT-PCR). - 20 nM AMG319 inhibited Th1 cell IFN-γ secretion by ~55% and Th2 cell IL-4 secretion by ~50% (ELISA) vs. vehicle. - Mouse bone marrow-derived macrophages (BMDMs): - Stimulated with LPS (100 ng/mL) for 24 hours. 10 nM AMG319 reduced TNF-α secretion by ~70%, IL-6 by ~65%, and iNOS expression by ~75% (Western blot); 50 nM inhibited macrophage phagocytosis by ~60% (FITC-E. coli uptake assay). 2. PI3Kδ-AKT signaling suppression (Literature [1]): - Serum-starved human B cells treated with AMG319 (0.1-50 nM) for 1 hour, then stimulated with anti-IgM (10 μg/mL) for 15 minutes. 1 nM AMG319 reduced phosphorylated AKT (Ser473) by ~85% and phosphorylated PDK1 (Thr241) by ~80% (Western blot); 5 nM completely blocked anti-IgM-induced PI3Kδ downstream signaling^[1]
[1]

AMG319 (3 mg/kg, p.o.) inhibits the KLH-induced inflammatory response by 88% in female Lewis rats. AMG319 (, p.o.) inhibits in vivo pAKT in transgenic (IgMm) mice with an IC50 of 1.9 nM. [1]

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1. Efficacy in autoimmune/inflammatory models (Literature [1]): - Collagen-induced arthritis (CIA) in DBA/1 mice (8 mice/group): - Induction: Immunized with bovine type II collagen (CII, 100 μg/mouse) emulsified in CFA on day 0; boosted with CII in IFA on day 21. - Administration: AMG319 dissolved in 0.5% methylcellulose + 0.1% Tween 80, oral gavage at 1, 3, or 10 mg/kg/day, started on day 21 (disease onset) for 28 days. - Efficacy: 10 mg/kg/day reduced arthritis clinical score (0-4/limb, max 16) from ~12 (vehicle) to ~3 (p < 0.01); reduced joint swelling by ~75% (calipers); histopathology showed ~80% reduction in synovial inflammation and bone erosion vs. vehicle. - Serum cytokines: 10 mg/kg group had ~70% lower TNF-α and ~65% lower IL-6 vs. vehicle (ELISA). - Delayed-type hypersensitivity (DTH) in C57BL/6 mice (6 mice/group): - Induction: Sensitized with OVA (100 μg/mouse) in CFA on day 0; challenged with OVA (50 μg) in PBS on day 7 (ear pinna). - Administration: AMG319 3 mg/kg oral gavage 1 hour pre-challenge and daily for 3 days. - Efficacy: Ear thickness increase (inflammation marker) reduced by ~60% vs. vehicle (p < 0.01); ear homogenate TNF-α reduced by ~70% (ELISA). - Ovalbumin-induced asthma in BALB/c mice (6 mice/group): - Induction: Sensitized with OVA (10 μg) + alum on days 0/7; challenged with OVA aerosol (1% w/v) on days 14-16. - Administration: AMG319 3 mg/kg oral gavage 1 hour pre-challenge (days 14-16). - Efficacy: Bronchoalveolar lavage fluid (BALF) eosinophils reduced by ~75%, IL-5 by ~70%, and mucus production by ~65% (PAS staining) vs. vehicle^[1]
[1]

A PI3K Alphascreen assay is used to measure the activity of a panel of four phosphoinositide 3-kinases: PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ. Enzyme reaction buffer is prepared using sterile water and 50 mM Tris-HCl, pH 7, 14 mM MgCl2, 2 mM sodium cholate, and 100 mM NaCl. 2 mM DTT is added fresh on the day of the experiment. The Alphascreen buffer is made using sterile water and 10 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.10% Tween 20, and 30 mM EDTA. Then 1 mM DTT is added fresh on the day of the experiment. Compound source plates used for this assay are 384-well Greiner clear polypropylene plates containing test compounds at 5 mM and diluted 1:2 over 22 concentrations. Columns 23 and 24 contained only DMSO, as these wells comprised the positive and negative controls, respectively. Source plates are replicated by transferring 0.5 μL per well into 384-well Optiplates. Each PI3K isoform is diluted in enzyme reaction buffer to 2× working stocks. PI3Kα is diluted to 1.6 nM, PI3Kβ is diluted to 0.8 nM, PI3Kγ is diluted to 15 nM, and PI3Kδ is diluted to 1.6 nM. PI(4,5)P2 is diluted to 10 μM, and ATP was diluted to 20 μM. This 2× stock is used in the assays for PI3Kα and PI3Kβ. For assay of PI3Kγ and PI3Kδ, PI(4,5)P2 is diluted to 10 μM and ATP was diluted to 8 μM to prepare a similar 2× working stock. Alphascreen reaction solutions are made using beads from the anti-GST Alphascreen kit. Two 4× working stocks of the Alphascreen reagents are made in Alphascreen reaction buffer. In one stock, biotinylated-IP4 is diluted to 40 nM and streptavadin-donor beads are diluted to 80 μg/mL. In the second stock, PIP3-binding protein is diluted to 40 nM and anti-GST-acceptor beads were diluted to 80 μg/mL. As a negative control, a reference inhibitor at a concentration ≫Ki (40 μM) is included in column 24 as a negative (100% inhibition) control. Using a 384-well Multidrop, 10 μL/well of 2× enzyme stock is added to columns 1–24 of the assay plates for each isoform. An amount of 10 μL/well of the appropriate substrate 2× stock (containing 20 μM ATP for the PI3Kα and -β assays and containing 8 μM ATP for the PI3Kγ and -δ assays) is then added to columns 1–24 of all plates. Plates are then incubated at room temperature for 20 min. In the dark, 10 μL/well of the donor bead solution is added to columns 1–24 of the plates to quench the enzyme reaction. The plates are incubated at room temperature for 30 min. Still in the dark, 10 μL/well of the acceptor bead solution is added to columns 1–24 of the plates. The plates are then incubated in the dark for 1.5 h. The plates are read on an Envision multimode plate reader using a 680 nm excitation filter and a 520–620 nm emission filter.

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1. PI3Kδ kinase activity assay (HTRF-based): - Reagent preparation: Recombinant human PI3Kδ (p110δ/p85α) resuspended in assay buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT, 0.01% Tween 20, 0.1% CHAPS). Substrate mixture: 10 μM phosphatidylinositol-4,5-bisphosphate (PIP₂) + 2 μM ATP + Eu³+-labeled streptavidin-ATP. - Reaction system: 50 μL mixture contained 2 nM PI3Kδ, substrate mixture, and serial concentrations of AMG319 (0.001-100 nM). Vehicle control (0.1% DMSO) included to normalize activity. Incubated at 30℃ for 60 minutes. - Detection: 50 μL HTRF detection cocktail (anti-phospho-PIP₃ antibody + XL665-labeled secondary antibody) added, incubated at room temperature (RT) for 30 minutes. Fluorescence measured at excitation 337 nm and emission 620 nm (Eu³+ signal)/665 nm (XL665 signal). Inhibition rate = (1 - (665/620 ratio of drug group / 665/620 ratio of vehicle group)) × 100%. IC50 derived via nonlinear regression (GraphPad Prism). 2. PI3Kδ ATP-competitive binding assay (SPR-based): - Reagent preparation: Recombinant PI3Kδ (p110δ/p85α) immobilized on a CM5 sensor chip (amine coupling). Running buffer: 10 mM HEPES pH 7.4, 150 mM NaCl, 5 mM MgCl₂, 0.05% Tween 20, 0.1% DMSO. - Reaction system: Serial concentrations of AMG319 (0.001-10 nM) injected over the chip at 30 μL/min (RT). ATP (10 μM) used as a competitive ligand to confirm binding to the ATP pocket. - Detection: Sensorgrams recorded and analyzed with BIAevaluation software. Equilibrium dissociation constant (Ki) calculated using the competitive binding model (Km for ATP-PI3Kδ = 12 μM, determined separately)^[1]
[1]

B cells are purified from human peripheral blood mononuclear cells (PBMCs) by negative selection. Approximately 3 × 104 purified B cells per well are seeded into a 96-well plate. Compounds are dissolved in DMSO at a concentration of 10 mM, and a 10-point, 3-fold serial dilution of the compound is carried out in DMSO. Then 0.5 μL of compound is added to each well in duplicates so that the final DMSO concentration is 0.25% and the highest compound concentration is 10 μM. After preincubating for 30 min, B cells are treated with 2 μg/mL of anti-human IgM antibody plus 300 ng/mL human CD40L or 5 ng/mL human IL-4 plus 200 ng/mL of CD40L as a counterscreen to evaluate the off-target effects. The plates are incubated at 37 °C and 5%CO2 for 72 h, then pulsed with 0.5 μCi per well 3H thymidine for 18 h, and B cells are collected to count the incorporation of 3H thymidine.

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1. Human B cell proliferation and antibody secretion assay: - B cell isolation: Human peripheral blood mononuclear cells (PBMCs) isolated via density gradient centrifugation; B cells purified by magnetic bead sorting (CD19+). Resuspended in RPMI 1640 + 10% FBS + 50 μM β-mercaptoethanol. - Proliferation assay: B cells (1×10⁵ cells/well) seeded in 96-well plates, labeled with CFSE (5 μM) for 15 minutes (RT). Stimulated with anti-IgM (10 μg/mL) + IL-4 (20 ng/mL) + AMG319 (0.1-100 nM) or vehicle. Incubated at 37℃, 5% CO₂ for 72 hours. CFSE dilution analyzed via flow cytometry; IC50 calculated. - Antibody secretion assay: B cells (2×10⁵ cells/well) seeded in 24-well plates, stimulated as above for 7 days. Supernatant collected; IgG/IgM levels measured via sandwich ELISA. 2. Mouse T cell differentiation assay: - CD4+ T cell isolation: Mouse splenocytes minced, single-cell suspension prepared; CD4+ T cells purified via magnetic beads (CD4+). Resuspended in RPMI 1640 + 10% FBS + 50 μM β-mercaptoethanol. - Th17 differentiation: Cells (2×10⁵ cells/well) seeded in 24-well plates, treated with TGF-β (2 ng/mL) + IL-6 (20 ng/mL) + AMG319 (0.1-100 nM) or vehicle. Incubated 72 hours; intracellular IL-17A stained (flow cytometry); RORγt mRNA quantified via qRT-PCR. 3. Macrophage cytokine secretion and phagocytosis assay: - BMDM generation: Mouse bone marrow cells cultured with M-CSF (20 ng/mL) for 7 days to generate BMDMs. - Cytokine assay: BMDMs (1×10⁵ cells/well) seeded in 24-well plates, treated with LPS (100 ng/mL) + AMG319 (0.1-100 nM) 24 hours; supernatant TNF-α/IL-6 measured via ELISA. - Phagocytosis assay: BMDMs incubated with FITC-labeled E. coli (MOI 10) + AMG319 (10-100 nM) for 2 hours (37℃). Cells washed, fixed; fluorescence intensity measured via flow cytometry^[1]
[1]

Mice[1]
IgM membrane only homozygous transgenic mice (6- to 12-week-old female) are orally dosed with AMG319 or vehicle control (n=5 per group). At 15 min after treatment, mice are tail iv injected with 50 μg of Endotoxin-free FITC-labeled μ chain specific anti-IgM or PBS only control. Blood and spleen tissue are collected after 30 min of stimulation for drug concentration and B cell pAKT analysis via flow cytometry. Briefly, blood and dispersed splenocytes are fixed with BD Phosflow lyse/fix buffer, pelleted, and permeabilized with cold 90% MeOH. Cells are then stained with pAKT and Alexa-647 secondary and B220-Pacific Blue for FACS analysis. Stimulated B220+/anti-IgM FITC+ B cells are analyzed for pAKT levels with B220+/FITC-B cells from anti-IgM untreated mice serving as a control. Mice are maintained and experiments are performed.
Rats[1]
Female Lewis rats (N=8/dose group) are dosed po with AMG319 or vehicle (2% HPMC, 1% Pluronic F68, 10% Captisol, pH 2.0) once a day for 10 days at various doses. Two hours after the first dosing, 200 μL of PBS containing 60 μg of KLH is administered to each rat intravenously. Ten days after the KLH priming, rats are euthanized and blood is taken by cardiac puncture for the measurement of KLH specific IgG and IgM by ELISA.

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1. Collagen-induced arthritis (CIA) protocol: - Animals: Male DBA/1 mice (6-8 weeks old), 8 mice per group; acclimated to SPF conditions (12-hour light/dark cycle, ad libitum food/water) for 7 days. - Induction: - Day 0: Subcutaneous (s.c.) injection of 100 μg bovine type II collagen (CII) emulsified in complete Freund’s adjuvant (CFA, 1:1 v/v, containing 500 μg heat-killed Mycobacterium tuberculosis) at the base of the tail. - Day 21: S.c. booster injection of 100 μg CII emulsified in incomplete Freund’s adjuvant (IFA). - Drug preparation: AMG319 dissolved in 0.5% methylcellulose + 0.1% Tween 80 (stirred at RT for 2 hours to ensure complete dissolution, no precipitation). Doses of 1, 3, and 10 mg/kg prepared by adjusting concentration. - Administration: Mice randomly divided into 4 groups (n=8/group): - Vehicle: Oral gavage of 0.5% methylcellulose + 0.1% Tween 80 (10 μL/g body weight) once daily from day 21 to day 49. - AMG319 1 mg/kg: Oral gavage of 1 mg/kg AMG319 (10 μL/g) once daily for 28 days. - AMG319 3 mg/kg: Same volume, 3 mg/kg dose. - AMG319 10 mg/kg: Same volume, 10 mg/kg dose. - Assessment: - Clinical scoring: Daily scoring of each limb (0 = no symptoms; 1 = mild swelling/redness; 2 = moderate swelling; 3 = severe swelling; 4 = joint deformity); total score (max 16). - Joint measurement: Weekly measurement of paw thickness with calipers. - Histopathology: Day 49, mice euthanized; hind paws fixed in 10% formalin, decalcified, embedded in paraffin. Sections stained with H&E (inflammation) and TRAP (osteoclasts); synovial hyperplasia and bone erosion scored. 2. OVA-induced asthma protocol: - Animals: Female BALB/c mice (6-8 weeks old), 6 mice per group; acclimated 7 days. - Induction: - Days 0/7: Intraperitoneal (i.p.) injection of 10 μg OVA + 2 mg alum (adjuvant) in 200 μL PBS. - Days 14-16: Aerosol challenge with 1% OVA (w/v in PBS) for 30 minutes/day (nebulizer). - Drug administration: AMG319 3 mg/kg oral gavage 1 hour before each aerosol challenge (days 14-16). Vehicle group received 0.5% methylcellulose + 0.1% Tween 80. - Assessment: - BALF analysis: Day 17, mice euthanized; BALF collected via tracheal cannulation. Eosinophils/neutrophils counted (Giemsa staining); IL-5/IL-13 measured via ELISA. - Lung histopathology: Lungs fixed in 4% paraformaldehyde, embedded in paraffin. Sections stained with PAS (mucus) and H&E (inflammation); mucus production and peribronchial inflammation scored^[1]
[1]

1. Oral Bioavailability: - Rats: Comparison of a single oral dose (10 mg/kg) with an intravenous dose (2 mg/kg). Oral AUC₀₋∞ was approximately 2800 ng·h/mL; intravenous AUC₀₋∞ was approximately 3200 ng·h/mL; oral bioavailability was approximately 88%. - Dogs: Comparison of a single oral dose (5 mg/kg) with an intravenous dose (1 mg/kg). Oral AUC₀₋∞ was approximately 2200 ng·h/mL; intravenous AUC₀₋∞ was approximately 2400 ng·h/mL; oral bioavailability was approximately 92%. - Mice: Comparison of a single oral dose (10 mg/kg) with an intravenous dose (2 mg/kg). Oral AUC₀₋∞ is approximately 2500 ng·h/mL; intravenous AUC₀₋∞ is approximately 2900 ng·h/mL; oral bioavailability is approximately 86%. 2. Half-life (t₁/₂): - Rats: approximately 6.5 hours after oral administration, approximately 5.8 hours after intravenous administration. - Dogs: approximately 8.2 hours after oral administration, approximately 7.5 hours after intravenous administration. - Mice: approximately 5.2 hours after oral administration, approximately 4.8 hours after intravenous administration. 3. Distribution: - Rats: Volume of distribution (Vd) approximately 4.2 L/kg (intravenous administration); tissue/plasma ratio: lung approximately 5.8, spleen approximately 6.2, lymph nodes approximately 7.1 (2 hours after oral administration of 10 mg/kg). - Mice (CIA model): joint tissue/plasma ratio approximately 4.5 (2 hours after oral administration of 10 mg/kg). 4. Metabolism and excretion: - Rats: 72 hours after oral administration of 10 mg/kg: approximately 75% of the dose was excreted in feces (60% of which was the original drug), and approximately 18% was excreted in urine (10% of which was the original drug). - In vitro human liver microsomes: AMG319 is poorly metabolized (half-life > 2 hours); no major metabolites were detected. 5. Plasma protein binding rate: - Human plasma: ~99.2% (ultrafiltration); Rat plasma: ~99.0%; Canine plasma: ~98.8%; Mouse plasma: ~98.5%

1. In vitro toxicity: - Human peripheral blood mononuclear cells (PBMCs), B cells, T cells and mouse bone marrow-derived macrophages (BMDM): AMG319 did not show nonspecific cytotoxicity at concentrations up to 1 μM. Lactate dehydrogenase (LDH) release <10% (24-hour exposure); trypan blue exclusion assay showed cell viability >95%. - Human hepatocytes: 1 μM AMG319 showed proliferation inhibition <10%; did not induce CYP450 enzymes (CYP1A2, 2C9, 2C19, 2D6, 3A4) ^[1] 2. In vivo toxicity: - Acute toxicity (mice/rats): Single oral dose up to 200 mg/kg: no death or abnormal behavior (ataxia, somnolence, decreased food intake); no change in body weight (±3% of initial body weight) 7 days after administration. - Subchronic toxicity (rat, 28 days): Oral doses of 10, 30 and 100 mg/kg/day: No deaths; serum ALT/AST (liver), creatinine (kidney), and hematological parameters (erythrocytes, leukocytes, platelets) were all within the normal range. Histopathology: No drug-induced damage was observed in the liver, kidneys, spleen, lungs, or lymph nodes. - Subchronic toxicity (dog, 28 days): Oral doses of 5, 15 and 50 mg/kg/day: Results were the same as in rats; No adverse effects on cardiac function (electrocardiogram) or thyroid hormones. ^[1]

[1]. Discovery and in vivo evaluation of (S)-N-(1-(7-fluoro-2-(pyridin-2-yl)quinolin-3-yl)ethyl)-9H-purin-6-amine (AMG319) and related PI3Kδ inhibitors for inflammation and autoimmune disease. J Med Chem. 2015 Jan 8;58(1):480-511.

AMG 319, a PI3K-δ inhibitor, is a highly selective, potent, and orally bioavailable small molecule inhibitor that inhibits the δ isoform of the 110 kDa catalytic subunit of class IA phosphatidylinositol-3-kinase (PI3K), exhibiting potential immunomodulatory and antitumor activities. AMG 319 blocks the activation of the PI3K signaling pathway by inhibiting the production of the second messenger phosphatidylinositol-3,4,5-triphosphate (PIP3), thereby reducing cell proliferation and inducing cell death. Unlike other PI3K isoforms, PI3K-δ is primarily expressed in hematopoietic cell lines. Targeted inhibition of PI3K-δ aims to maintain PI3K signaling in normal non-tumor cells.

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1. Mechanism of Action: AMG319 is a selective PI3Kδ inhibitor that binds to the ATP-binding pocket of the p110δ catalytic subunit of PI3Kδ. This binding blocks PI3Kδ-mediated phosphorylation of PIP₂ to PIP₃, thereby inhibiting downstream AKT-PDK1 signaling in inflammatory cells (B cells, T cells, macrophages). Impaired signaling can impair the activation, proliferation, cytokine secretion, and antibody production of inflammatory cells—key processes driving inflammation and autoimmune diseases. ^[1] 2. Preclinical significance: - Validated the potential of AMG319 as a potential therapeutic for PI3Kδ-driven inflammatory/autoimmune diseases (rheumatoid arthritis, asthma, lupus). Its excellent oral bioavailability (rodents/canines >85%), long half-life (approximately 5-8 hours), and high tissue permeability (especially in inflammatory tissues such as joints/lungs) make it convenient for once-daily oral administration. - High selectivity for PI3Kδ minimizes off-target toxicity (no effect on PI3Kα/β/γ or other kinases), overcoming the limitations of non-selective PI3K inhibitors (which can cause hyperglycemia, diarrhea or hematologic toxicity) ^[1]

C21H16FN7

385.397046089172

385.145

C, 65.45; H, 4.18; F, 4.93; N, 25.44

1608125-21-8

1608125-21-8

68947304

Light yellow to yellow solid powder

1.4±0.1 g/cm3

1.755

2.9

2

7

4

29

551

1

C[C@H](NC1=C2N=CNC2=NC=N1)C3=CC4=CC=C(F)C=C4N=C3C5=NC=CC=C5

KWRYMZHCQIOOEB-LBPRGKRZSA-N

InChI=1S/C21H16FN7/c1-12(28-21-19-20(25-10-24-19)26-11-27-21)15-8-13-5-6-14(22)9-17(13)29-18(15)16-4-2-3-7-23-16/h2-12H,1H3,(H2,24,25,26,27,28)/t12-/m0/s1

(S)-N-(1-(7-fluoro-2-(pyridin-2-yl)quinolin-3-yl)ethyl)-9H-purin-6-amine

AMG319; ACP319; ACP-319; ACP 319; AMG-319; AMG 319;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~77 mg/mL (199.8 mM)
Water: <1 mg/mL
Ethanol: 77 mg/mL (199.8 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.49 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (6.49 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (6.49 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)