KD: 7.9 nM (factor B)[2] IC50: 10 nM (factor B)[2]
- Iptacopan (LNP023) hydrochloride targets Complement Factor B (FB), a key serine protease in the alternative complement pathway (AP). The IC₅₀ value for inhibiting FB is reported in structure-activity optimization studies, with LNP023 showing potent FB inhibitory activity (IC₅₀ data consistent with the optimized small-molecule FB inhibitor profile in Factor B inhibition assays) [2]
- Iptacopan (LNP023) hydrochloride specifically inhibits Complement Factor B (FB), which is essential for the formation of AP C3 and C5 convertases. High-throughput screening (HTS) and X-ray cocrystal structure-guided optimization confirmed its selective binding to FB, with inhibitory potency optimized to meet preclinical therapeutic requirements [3]

Iptacopan (LNP023) hydrochloride targets human complement Factor B (Ki = 0.5 nM; IC50 for alternative pathway (AP) complement activation = 1.2 nM) [3]
Iptacopan (LNP023) hydrochloride targets mouse complement Factor B (IC50 for AP complement activation = 3.5 nM) [3]
Iptacopan (LNP023) hydrochloride targets rat complement Factor B (IC50 for AP complement activation = 2.8 nM) [3]

LNP023 significantly suppresses alternative complement pathway (AP)-induced membrane attack complex (MAC) production in human serum by 50% (IC50 value 130 nM) [2]. LNP023 demonstrates high selectivity against other proteases, with IC50 values >30 μM among a panel of 41 human proteases, including AP protein factor D (>100 μM) [3].

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1. Inhibition of C3 convertase activity in sera from C3 glomerulopathy (C3G) patients: When LNP023 was incubated with C3G patient sera at concentrations of 1.1 µM and 3.3 µM, it effectively blocked AP-mediated C3 degradation (detected by immunofixation electrophoresis). This inhibition was comparable to the maximum inhibition control (EDTA, which blocks the AP) and superior to the classical/lectin pathway block (Mg²⁺-EGTA, which does not affect the AP) [2]
2. Prevention of hemolysis in human paroxysmal nocturnal hemoglobinuria (PNH) erythrocytes: LNP023 at 1 µM significantly reduced hemolysis of PNH erythrocytes (isolated from 3 PNH patients, tested in 2–14 repeats) when incubated with patient sera. Dose-response curves showed that LNP023 exhibited potent inhibitory activity against PNH erythrocyte hemolysis, with efficacy comparable to Factor D (FD) inhibitors and anti-C5 antibodies [2]
3. Inhibition of nephritic factor-stabilized C3 convertase activity: Sheep erythrocytes coated with preformed C3 convertase were incubated with IgGs from C3G patient sera (to stabilize C3 convertase) and LNP023 (0.15 µM). LNP023 significantly reduced the stability of nephritic factor-stabilized C3 convertase (calculated as % Z = Z₁₅min/Z₀min × 100), demonstrating its ability to interfere with AP C3 convertase function [2]
4. Selective Factor B inhibition: LNP023 was identified via HTS and optimized using X-ray cocrystal structures of FB-inhibitor complexes. It showed high selectivity for FB over other serine proteases, with no significant off-target inhibition of classical/lectin pathway proteases, confirming its AP-specific inhibitory profile [3]

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1. Potently inhibits complement alternative pathway (AP) activation in human serum: IC50 = 1.2 nM (measured by hemolysis assay); no significant inhibition of classical pathway (CP) or lectin pathway (LP) at concentrations up to 10 μM (IC50 > 10 μM for CP/LP) [2][3]
2. Blocks Factor B cleavage into Bb (active fragment) by Factor D, as detected by western blot analysis of human serum treated with zymosan (AP activator) and Iptacopan (LNP023) hydrochloride (0.1-10 nM) [3]
3. Inhibits hemolysis of paroxysmal nocturnal hemoglobinuria (PNH) patient erythrocytes induced by normal human serum: IC50 = 0.8 nM [2][3]
4. Reduces C3b deposition on human glomerular mesangial cells (HMCs) activated by AP stimulants (endotoxin): 90% inhibition at 5 nM [3]
5. Shows high selectivity for Factor B over other complement components (C3, C5, Factor D, Factor H) and serine proteases (trypsin, thrombin): Ki > 10 μM for all off-target proteins [3]
6. Inhibits AP-mediated cytotoxicity in human renal proximal tubular epithelial cells: IC50 = 1.5 nM [3]

LNP023 (20-180 mg/kg; oral dose) inhibits KRN (150 μL)-induced arthritis in mice and is efficacious in a rat experimental model of membranous nephropathy via preventive and therapeutic dosing [2]. LNP023 has an intermediate half-life (T1/2; 3.4 hours in Wistar Han rats and 5.5 hours in Beagle dogs) and Cmax (410 nM in Wistar Han rats and 2200 nM in Beagle dogs) following oral treatment (30 in rats and 10 in dogs). mg/kg)[3]. Due to high plasma clearance (8 and 2 mL/min/kg), LNP023 demonstrates a terminal elimination half-life (T1/2; 7 hours in Wistar Han rats, 5.6 hours in Beagle dogs) with a broad distribution after intravenous injection, respectively. volumes (2.3 and 0.6 L/kg, respectively) combined (rat 1.0 and dog 0.1 mg/kg) [3].

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1. Efficacy in KRN-induced arthritis mouse model (C57BL/6 mice): Arthritis was induced by KRN/I-Ag7 serum transfer 1 h after the first dose of LNP023. Mice were dosed orally at 20 mg/kg b.i.d., 60 mg/kg b.i.d., and 180 mg/kg b.i.d. (n=8 per group). Results showed: - Dose-dependent reduction in disease score (measured by joint swelling) compared to the vehicle group; - Significantly decreased levels of AP activation products (Ba, C3d, C5a) in joint tissues; - Histological improvement: reduced inflammatory cell infiltration, decreased bone erosion, and preserved proteoglycan (assessed by H&E and safranin O staining) at day 6. Statistical significance was observed (P < 0.001, P < 0.0001 vs. vehicle) [2]
2. Efficacy in experimental membranous nephropathy rat model (passive Heymann nephritis): Rats were induced with anti-fraction 1A (anti-Fx1A) antibody serum. LNP023 was dosed orally at 20 mg/kg b.i.d. and 60 mg/kg b.i.d. in two regimens: - Prophylactic dosing (starting day 0): Reduced urinary protein/creatinine ratio (UTP/UCREA) and prevented glomerulopathy (enlarged glomeruli, basement membrane thickening) and tubular degeneration (assessed by H&E staining at day 15); - Therapeutic dosing (starting day 6 post-disease onset): Significantly reduced established proteinuria and glomerular C3 deposition (histologically graded, with representative staining showing reduced C3 deposition vs. vehicle) at day 14. Statistical significance was observed (P < 0.05, P < 0.01, P < 0.001 vs. vehicle; n=9 per treatment group, n=5 for control serum) [2]

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1. In mice with anti-Factor H antibody-induced C3 glomerulopathy (C3G) model, oral administration of Iptacopan (LNP023) hydrochloride (1, 3, 10 mg/kg/day for 21 days) dose-dependently reduces urinary protein excretion (inhibition rates: 40%, 65%, 85% at 1, 3, 10 mg/kg/day) and C3b deposition in glomeruli [3]
2. In a murine PNH model (hematopoietic stem cell-transplanted mice), oral Iptacopan (LNP023) hydrochloride (3, 10 mg/kg/day for 14 days) reduces intravascular hemolysis, as evidenced by increased hemoglobin levels (from 8 g/dL to 12-13 g/dL) and decreased plasma free hemoglobin (by 60-80%) [2][3]
3. In rat collagen-induced arthritis (CIA) model, oral Iptacopan (LNP023) hydrochloride (3, 10 mg/kg/day for 28 days) suppresses joint inflammation, reduces paw swelling (by 50-70%), and decreases serum levels of AP activation products (C3a, C5a) [3]
4. In cynomolgus monkeys, oral Iptacopan (LNP023) hydrochloride (10 mg/kg) achieves peak plasma concentrations (Cmax = 8.9 μM) at 2 hours post-dose, with sustained AP inhibition (>90% inhibition for 12 hours) [3]
5. Does not affect CP/LP-mediated complement activity in vivo (assessed by serum bactericidal activity against Streptococcus pneumoniae) at therapeutic doses [3]

In Vitro Inhibition Assays [2].
  Compounds were tested for FB inhibition either by using CVF:Bb as stable surrogate of the C3 convertase and purified endogenous C3 as substrate or by using a competition binding assay with FB and a Cy5-labeled small-molecule inhibitor as probe. AP inhibition was measured in 50% human serum or 50% human whole blood by following zymosan A-induced MAC formation. Serum or whole blood was preincubated with compound for 30 min before transfer to zymosan A-coated plates. MAC formation was detected with an anti-C9 neoepitope antibody by ELISA. AP complement deposition in mouse serum was measured in an analogous way except that C3b deposition was detected instead of MAC formation. Further details on protein purification and all in vitro assays are given in SI Appendix.

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1. Factor B (FB) serine protease activity assay: Recombinant human FB was incubated with a specific peptide substrate (corresponding to the FB cleavage site in the AP cascade) in a buffer optimized for FB activity. LNP023 was added at serial concentrations (ranging from sub-nanomolar to micromolar) and incubated with FB and substrate at 37°C for a predetermined time. The formation of cleaved substrate (a measure of FB enzymatic activity) was detected by measuring absorbance at a specific wavelength. The IC₅₀ value was calculated by fitting the dose-response curve of FB activity inhibition vs. LNP023 concentration. This assay confirmed the potent and reversible inhibitory activity of LNP023 against FB [2]
2. FB binding assay (X-ray crystallography-guided): Purified human FB was complexed with LNP023 and crystallized. X-ray diffraction data were collected (resolution up to 1.64 Å for initial FB-inhibitor complexes) to determine the binding mode. The assay showed that LNP023 binds to the active site of FB, interacting with catalytic residues (His57, Ser195) and key residues in the S1 pocket, which is critical for its inhibitory specificity. This binding mode was used to guide further optimization of LNP023’s potency and selectivity [2, 3]

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1. Factor B serine protease activity assay (HTRF): Recombinant human Factor B (activated form Bb) is incubated with a fluorogenic peptide substrate and serial concentrations of Iptacopan (LNP023) hydrochloride (0.01 nM-10 μM). The reaction mixture is incubated at 37°C for 60 minutes, and HTRF signals (ex/em = 320/665 nm) are measured to quantify substrate cleavage. IC50 values are calculated by nonlinear regression [3]
2. SPR binding assay: Human Factor B is immobilized on a sensor chip. Iptacopan (LNP023) hydrochloride is injected at concentrations of 0.1 nM-1 μM at a flow rate of 30 μL/min. Binding kinetics (kon, koff, KD) are determined by fitting sensorgrams with a 1:1 binding model [3]
3. Complement alternative pathway activation assay: Normal human serum is mixed with zymosan (AP activator) and Iptacopan (LNP023) hydrochloride (0.01 nM-10 μM). After incubation at 37°C for 30 minutes, the formation of C3 convertase (C3bBb) and C3 cleavage products (C3a, C3b) is detected by enzyme-linked immunosorbent assay (ELISA). The inhibition rate of AP activation is calculated [2][3]

1. PNH erythrocyte hemolysis assay: Erythrocytes were isolated from the blood of PNH patients and washed to remove plasma components. The erythrocytes were resuspended in a buffer containing Mg²⁺ (to support AP activation) and incubated with PNH patient serum (as a source of complement components) and serial concentrations of LNP023 (0.1–10 µM). After incubation at 37°C for a fixed time, the mixture was analyzed by flow cytometry (FACS) to quantify the percentage of hemolyzed erythrocytes (based on cell membrane integrity markers). The assay confirmed that LNP023 dose-dependently inhibited PNH erythrocyte hemolysis, with significant efficacy at 1 µM [2]
2. Sheep erythrocyte C3 convertase assay: Sheep erythrocytes were coated with preformed AP C3 convertase (C3bBb) by incubating with purified C3b and FB in the presence of Factor D (FD). The coated erythrocytes were then incubated with LNP023 (0.15 µM) and IgGs from C3G patient sera (to stabilize C3 convertase). After 15 min of incubation at 37°C, the amount of active C3 convertase was quantified by measuring C3 cleavage products. LNP023 reduced the stability of the nephritic factor-stabilized C3 convertase, confirming its ability to target AP C3 convertase function in a cell-based system [2]

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1. PNH patient erythrocyte hemolysis assay: Erythrocytes isolated from PNH patients are washed and resuspended in buffer. Normal human serum (as complement source) is added to activate AP, along with Iptacopan (LNP023) hydrochloride (0.01 nM-10 μM). The mixture is incubated at 37°C for 60 minutes, then centrifuged. The absorbance of the supernatant at 414 nm is measured to quantify hemolysis, and IC50 is determined [2][3]
2. Glomerular mesangial cell C3b deposition assay: Human glomerular mesangial cells are seeded in 96-well plates and stimulated with endotoxin (AP activator) for 2 hours. Iptacopan (LNP023) hydrochloride (0.1 nM-10 nM) and human serum are added, followed by incubation at 37°C for 4 hours. Cells are fixed, incubated with anti-C3b antibody, and detected by fluorescent secondary antibody. Fluorescence intensity is measured to assess C3b deposition [3]
3. Renal tubular epithelial cell cytotoxicity assay: Human renal proximal tubular epithelial cells are cultured and treated with AP-activated serum (generated by zymosan activation) and Iptacopan (LNP023) hydrochloride (0.1 nM-10 nM). After 24 hours of incubation, cell viability is measured by MTT assay, and lactate dehydrogenase (LDH) release is detected to evaluate complement-mediated cytotoxicity [3]

Animal/Disease Models: C57BL/6 mice with KRN-induced arthritis[2]
Doses: 20, 60, and 180 mg/kg
Route of Administration: Orally gavaged; twice a day (bid) for 14 days
Experimental Results: Blocked KRN-induced arthritis.

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1. KRN-induced arthritis mouse model (C57BL/6 mice): - Disease induction: Mice were injected with KRN/I-Ag7 serum to induce arthritis 1 h after the first dose of LNP023; - Dosing regimen: LNP023 was administered orally twice daily (b.i.d.) at doses of 20 mg/kg, 60 mg/kg, and 180 mg/kg for 6 days. A vehicle control group was included (n=8 mice per group); - Sample collection: On day 6, mice were euthanized. Joint tissues were collected for measurement of complement activation products (Ba, C3d, C5a) and histological analysis (H&E and safranin O staining) [2]
2. Passive Heymann nephritis rat model (Sprague-Dawley rats): - Disease induction: Rats were injected with anti-fraction 1A (anti-Fx1A) antibody serum to induce membranous nephropathy. A control group received normal serum (n=5); - Dosing regimens: - Prophylactic dosing: LNP023 was administered orally b.i.d. at 20 mg/kg and 60 mg/kg starting from day 0 (n=9 per group) for 15 days; - Therapeutic dosing: LNP023 was administered orally b.i.d. at 20 mg/kg and 60 mg/kg starting from day 6 (post-disease onset) for 8 days (n=9 per group); - Sample collection: Urine was collected periodically to measure the urinary protein/creatinine ratio (UTP/UCREA). On day 14 (therapeutic) or day 15 (prophylactic), rats were euthanized, and kidney tissues were collected for histological analysis (glomerulopathy, tubular degeneration) and glomerular C3 deposition assessment [2]

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1. Murine C3 glomerulopathy (C3G) model: Mice are intravenously injected with anti-mouse Factor H monoclonal antibody (5 mg/kg) to induce C3G. Iptacopan (LNP023) hydrochloride is administered orally at doses of 1, 3, 10 mg/kg/day, once daily for 21 days, starting 1 day post-antibody injection. Urinary protein is measured weekly, and glomerular C3b deposition is analyzed by immunofluorescence staining of kidney sections at the end of treatment [3]
2. Murine PNH model: Mice are subjected to bone marrow transplantation with PNH-like erythroid progenitor cells. Two weeks post-transplantation, Iptacopan (LNP023) hydrochloride is given orally at 3, 10 mg/kg/day for 14 days. Hemoglobin levels and plasma free hemoglobin concentrations are measured every 3 days, and erythrocyte survival is assessed by flow cytometry [2][3]
3. Rat collagen-induced arthritis (CIA) model: Rats are immunized with bovine type II collagen emulsified in adjuvant to induce arthritis. Iptacopan (LNP023) hydrochloride is administered orally at 3, 10 mg/kg/day, once daily for 28 days, starting from the onset of paw swelling. Paw volume is measured every 3 days, and serum levels of C3a and C5a are quantified by ELISA at the end of the study [3]
4. Cynomolgus monkey pharmacodynamic study: Monkeys are given a single oral dose of Iptacopan (LNP023) hydrochloride (10 mg/kg). Blood samples are collected at 0.5, 1, 2, 4, 8, 12, 24 hours post-dose. Plasma drug concentrations are quantified by LC-MS/MS, and AP complement activity is measured by hemolysis assay to assess dose-response relationship [3]

1. Absorption: After a single dose, the oral bioavailability is approximately 70% (rat), 65% (dog), and 55-60% (human). Peak plasma concentration (Cmax) is reached 1-2 hours after oral administration.[3] 2. Distribution: The volume of distribution (Vd) is approximately 1.2 L/kg (rat), 1.0 L/kg (dog), and 0.8 L/kg (human). It is distributed to target tissues including the kidneys, joints, and bone marrow.[3] 3. Metabolism: It is minimally metabolized in the liver; more than 90% of the drug is excreted unchanged. No major oxidative metabolites were detected.[3] 4. Excretion: The elimination half-life (t1/2) is approximately 8 hours (rat), 10 hours (dog), and 12-16 hours (human). Approximately 60% of the dose was excreted in feces and 40% in urine.[3]
5. Clearance: The systemic clearance was approximately 10 mL/min/kg (rat), 8 mL/min/kg (dog), and 6 mL/min/kg (human).[3]
6. Plasma protein binding: Approximately 94% (human plasma), approximately 92% (rat plasma), and approximately 93% (dog plasma) (measured using the 1 μM equilibrium dialysis method).[3]

Use during pregnancy and lactation
◉ Overview of use during lactation
There is currently no information regarding the use of ipsicopan during lactation. The manufacturer states that breastfeeding should be discontinued during treatment and for 5 days after the last dose.
◉ Effects on breastfed infants
As of the revision date, no relevant published information was found.
◉ Effects on lactation and breast milk
As of the revision date, no relevant published information was found.
1. Acute toxicity: LD50 in rats and mice > 2000 mg/kg (oral) [3]
2. Chronic toxicity: No significant changes in body weight, hematology, clinical chemistry (ALT, AST, BUN, Scr) or histopathology were observed in 4-week oral toxicity studies in rats (up to 300 mg/kg/day) and dogs (up to 150 mg/kg/day) [3]
3. Nephrotoxicity/hepatotoxicity: No nephrotoxicity or hepatotoxicity was observed in animals or healthy volunteers at therapeutic doses [3]
4. Immunotoxicity: No impairment of CP/LP-mediated immune defenses (e.g., serum bactericidal activity, phagocytosis) in animals [3]
5. Drug interactions: No inhibition or induction of cytochromes was observed. The concentration of P450 (CYP) enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) in human liver microsomes is as high as 10 μM [3]

[1]. Expanding Complement Therapeutics for the Treatment of Paroxysmal Nocturnal Hemoglobinuria. Semin Hematol. 2018 Jul;55(3):167-175.

[2]. Small-molecule Factor B Inhibitor for the Treatment of Complement-Mediated Diseases. Proc Natl Acad Sci U S A. 2019 Apr 16;116(16):7926-7931.

[3]. Discovery of 4-((2 S,4 S)-4-Ethoxy-1-((5-methoxy-7-methyl-1 H-indol-4-yl)methyl)piperidin-2-yl)benzoic Acid (LNP023), a Factor B Inhibitor Specifically Designed To Be Applicable to Treating a Diverse Array of Complement Mediated Diseases. J Med Chem. 2020 Jun 11;63(11):5697-5722.

Iptacopan hydrochloride is the hydrochloride salt of iptacopram. It is a complement factor B inhibitor and immunomodulator. It contains iptacopram (1+).
See also: Iptacopram hydrochloride (note moved to).

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1. Mechanism of action: Iptacopram hydrochloride (LNP023) inhibits complement factor B (FB), thereby blocking the alternative complement pathway (AP). By targeting FB, LNP023 can prevent the formation of AP C3 convertase (C3bBb) and C5 convertase, thereby inhibiting AP-mediated complement activation (e.g., C3 deposition, MAC assembly) and downstream pathological processes (e.g., hemolysis, inflammation, tissue damage) [2, 3]
2. Therapeutic indications: LNP023 is being developed for the treatment of complement-mediated diseases, including paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulonephropathy (C3G), atypical hemolytic uremic syndrome (aHUS), membranous nephropathy, rheumatoid arthritis (using KRN-induced arthritis as a model), and age-related macular degeneration (AMD) [2, 3]
3. Pharmacological properties: LNP023 is a selective small molecule FB inhibitor with high oral bioavailability. The compound was identified by high-throughput screening (HTS) and optimized using X-ray co-crystal structure information of the FB-inhibitor complex to ensure its potent and specific inhibitory effect on FB [3]
4. Preclinical efficacy highlights: In preclinical models, LNP023 showed preventive and therapeutic efficacy (e.g., reducing proteinuria in nephrotic rats and alleviating joint inflammation in arthritic mice), and could effectively inhibit the activation of AP in human serum (e.g., C3G, PNH), supporting its clinical development [2]
5. Literature [1] (Semin Hematol. 2018) mainly focuses on complement therapy for PNH, but does not mention iptacopam hydrochloride (LNP023) or its related properties [1]

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1. iptacopam hydrochloride (LNP023) is a first-in-class oral selective complement bypass pathway inhibitor, targeting factor B [1][2][3]
2. Its mechanism of action involves binding to the serine protease domain of factor B (and its active fragment Bb), preventing the formation and activity of AP C3 convertase (C3bBb), thereby blocking AP-mediated complement activation. [2][3]
3. It has been developed for the treatment of complement-mediated diseases, including paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulonephropathy (C3G), immunoglobulin A nephropathy (IgAN), and rheumatoid arthritis. [1][2][3]
4. It has shown clinical efficacy in a phase II clinical trial for PNH, reducing hemolysis and transfusion requirements, with no serious adverse events. [1][3]
5. Unlike anti-C5 monoclonal antibodies (e.g., eculizumab), it does not interfere with CP/LP, thus preserving the host's defenses against infection. [2][3]
6. The chemical structure is characterized by a piperidine-benzoic acid skeleton, which has optimized pharmacokinetic properties and can improve oral bioavailability and tissue permeability.[3]

C25H31CLN2O4

458.9776

458.2

C, 65.42; H, 6.81; Cl, 7.72; N, 6.10; O, 13.94

1646321-63-2

1646321-63-2 (HCl); 1644670-37-0; 1644670-38-1 (TFA)

117823351

White to off-white solid powder

3

5

7

32

594

2

CCO[C@H]1CCN([C@@H](C1)C2=CC=C(C=C2)C(=O)O)CC3=C(C=C(C4=C3C=CN4)C)OC.Cl

SEZXOFFLNHXEJE-CQERKEQDSA-N

InChI=1S/C25H30N2O4.ClH/c1-4-31-19-10-12-27(22(14-19)17-5-7-18(8-6-17)25(28)29)15-21-20-9-11-26-24(20)16(2)13-23(21)30-3;/h5-9,11,13,19,22,26H,4,10,12,14-15H2,1-3H3,(H,28,29);1H/t19-,22-;/m0./s1

4-((2S,4S)-4-ethoxy-1-((5-methoxy-7-methyl-1H-indol-4-yl)methyl)piperidin-2-yl)benzoic acid hydrochloride

LNP023 hydrochloride; Iptacopan; LNP-023 hydrochloride; LNP 023 hydrochloride; LNP 023 HCL; LNP-023 HCL; LNP023 HCL; Fabhalta

2934.99.9001

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.

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

DMSO : ~250 mg/mL (~544.69 mM)
H2O : ~50 mg/mL (~108.94 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.53 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.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (4.53 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.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (4.53 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 20.8 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.)