GT1a(EC50=34 pM);GT1b(EC50=4 pM)

Ledipasvir (also known as GS5885) is a HCV NS5A polymerase inhibitor that is used for the treatment of hepatitis C virus infection. The combination product of ledipasvir 90 mg/sofosbuvir 400 mg (trade name Harvoni) was approved by FDA in October 2014. The ledipasvir/sofosbuvir combination is a direct-acting antiviral agent that interferes with HCV replication and can be used to treat patients with genotypes 1a or 1b without PEG-interferon or ribavirin. Ledipasvir has an extended plasma half-life of 37-45 h in healthy volunteers and produces a rapid >3 log viral load reduction in monotherapy at oral doses of 3 mg or greater with once-daily dosing in genotype 1a HCV-infected patients. It has been shown to be safe and efficacious, with SVR12 rates up to 100% when used in combination with direct-acting antivirals having complementary mechanisms.

In clinical trials, it was observed ledpasvir was well tolerated and exhibited median maximal reduction of HCV RNA ranging from 2.3 log10 IU/ml to 3.3 log10 IU/ml. Emax modeling also showed administration of 30 mg ledpasvir after 3 days resulted in >95% maximal response of HCV RNA reduction to genotype 1a.Finally, it was also observed that HCV RNA was more sustained in genotype 1b compared to 1a.

Competitive Protein Binding Assay[1]
Human plasma and cell-culture medium containing 10% fetal bovine serum (CCM) were spiked with the test compound at a final concentration of 2 μM. Spiked plasma (1 mL) and CCM (1 mL) were placed into opposite sides of the assembled dialysis cells, which are separated by a semipermeable membrane. The dialysis cells were rotated slowly in a 37 °C water bath for the time necessary to reach equilibrium. Postdialysis plasma and CCM weights were measured, and the test compound concentrations in plasma and CCM were determined with LC/MS/MS.

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Metabolic Stability[1]
Metabolic stability in vitro was determined using pooled hepatic microsomal fractions (final protein concentration of 0.5 mg/mL) at a final test compound concentration of 3 μM. The reaction was initiated by the addition of an NADPH-regenerating system. Aliquot of 25 μL of the reaction mixture were transferred at various time points to plates containing a quenching solution. The test compound concentration in the reaction mixture was determined with LC/MS/MS. Hepatic intrinsic clearance was calculated as described previously by Obach, and the predicted clearance was calculated using the well-stirred liver model without protein restriction.
Metabolic stability was also determined in cryopreserved hepatocytes using tritiated test compounds. The incubation mixture contained 1 × 106 hepatocytes/mL and 1 μM tritiated test compound (2.5 μCi). The incubation was carried out with gentle shaking at 37 °C under a humid atmosphere of 95% air/5% CO2 (v/v). Aliquots of 50 μL were removed after 0, 1, 3, and 6 h and added to 100 μL of quenching solution. The samples were analyzed on a flow scintillation radio detector coupled to an HPLC system. The metabolites were quantified on the basis of the peak areas from the radio detector, with the cell-free control samples used as a reference. Metabolic stabilities in hepatocytes were determined by measuring the rate of disappearance of the test compound as the percent of total peak areas of the formed radiolabeled metabolites and the test compound.

Ledipasvir is a specific inhibitor of HCV NS5A protein to inhibit HCV replication in the HCV subgenomic replicon system. NS5A replication complex inhibitors are novel antiviral factors for HCV treatment. Typically, these inhibitors have high efficiency and low viral resistance when compared to traditional HCV replication inhibitor targeted on NS3 helicase and NS5B RNA polymerasae. NS5A inhibitors are supposed to bind across the NS5A dimer interface, proximal to N-terminal domain 1. The binding is thought to distort dimer association directly or allosterically, which may disrupt NS5A function in HCV RNA replication. When a JFH1/3a-NS5A hybrid replicon was used to assess susceptibility to NS5A, another inhibitor DCV was shown to be more potent than ledipasvir. Additionally, NS5A-A30K and -Y93H variants exhibited reduced sensitivity to ledpasvir (EC50 value of 1770 nM and 4300 nM respectively).

Absorption, Distribution, and Excretion
Absorption
After oral administration, ledipasvir reaches peak plasma concentrations in approximately 4 to 4.5 hours, with a peak concentration (Cmax) of 323 ng/mL.
Excretion Routes
Following a single oral dose of 90 mg [14C]-ledipasvir, the average total recovery of the [14C]-radioactive substance in feces and urine is approximately 87%, with the majority of the radioactive dose recovered in feces (approximately 86%). On average, 70% of the administered dose of unchanged ledipasvir is excreted in feces, and 2.2% is excreted as the oxidative metabolite M19. These data suggest that bile excretion is the primary route of excretion for unchanged ledipasvir, while renal excretion is a secondary route (approximately 1%). Metabolites/Metabolites
In vitro studies did not detect metabolism of ledipasvir by human CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4. Evidence of slow oxidative metabolism via an unknown mechanism has been observed. Following a single 90 mg [14C]-ledipasvir dose, systemic exposure is almost entirely unchanged (>98%). Unmetabolized ledipasvir is predominantly present in feces.
Biological Half-Life
The median terminal half-life of ledipasvir is 47 hours.

Effects during pregnancy and lactation
◉ Summary of medication use during lactation
Studies of ledipasvir in breastfeeding women receiving treatment for hepatitis C virus infection have not been conducted. Because it binds to maternal plasma proteins at a rate as high as 99.8%, its concentration in breast milk may be very low. Breastfeeding does not need to be discontinued if the mother is using ledipasvir alone or in combination with sofosbuvir (Harvoni). Some sources suggest that breastfeeding should be avoided when ledipasvir is used in combination with ribavirin.
Hepatitis C is not transmitted through breast milk, and breast milk has been shown to inactivate the hepatitis C virus (HCV). However, the U.S. Centers for Disease Control and Prevention (CDC) recommends that breastfeeding should be considered if a mother infected with hepatitis C experiences nipple fissures or bleeding. It is currently unclear whether this warning applies to mothers receiving treatment for hepatitis C. Infants born to mothers infected with hepatitis C virus (HCV) should be tested for HCV; nucleic acid testing is recommended because maternal antibodies are present in the infant during the first 18 months of life and before the infant develops an immune response. ◉ Effects on breastfed infants
No published information found as of the revision date.
◉ Effects on breastfeeding and breast milk
No published information found as of the revision date.
Drug and Lactation Database (LactMed)
Protein binding rate
Leadipasvir binds to human plasma proteins >99.8%.

[1]. Discovery of ledipasvir (GS-5885): a potent, once-daily oral NS5A inhibitor for the treatment of hepatitis C virus infection. J Med Chem. 2014 Mar 13;57(5):2033-46

[2]. Natural prevalence of NS5A polymorphisms in subjects infected with hepatitis C virus genotype 3 and their effects on the antiviral activity of NS5A inhibitors. J Clin Virol. 2013 May;57(1):13-8.

We have discovered a new class of highly effective NS5A inhibitors with an asymmetric benzimidazole-difluorofluorene-imidazolium ring system at the core and a [2.2.1] azabicyclic ring system at the distal end. By optimizing antiviral activity and pharmacokinetics, we screened compound 39 (ledipasvir, GS-5885). Compound 39 (GT1a replicon EC50 = 31 pM) has a plasma half-life of 37-45 hours in healthy volunteers and rapidly reduces viral load by more than 3 logs in patients with genotype 1a HCV infection with a single oral dose of 3 mg or higher once daily. Studies have shown that compound 39 is safe and effective, and when used in combination with direct-acting antiviral drugs with complementary mechanisms of action, the SVR12 rate is as high as 100%. [1]

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Background: In vitro studies have shown that hepatitis C virus (HCV) NS5A replication complex inhibitors (RCIs) have picomolar-level antiviral activity against genotype 1 (GT1). This translates to a rapid and significant decrease in HCV RNA levels in GT1 patients. However, little is known about the sensitivity of other genotypes (e.g., GT3) to NS5A RCI inhibition. Objective: To detect and analyze the phenotypes of naturally occurring HCV GT3 NS5A polymorphisms in two currently clinically developed NS5A RCIs (daclatasvir [DCV] and GS-5885). Study Design: The NS5A regions of 96 treatment-naïve HCV GT3 patients from North America, Europe, and Australia were analyzed. Results: Phylogenetic analysis showed a wide distribution with no significant geographic clustering. GT1 DCV resistance-associated variants (RAVs) were observed in GT3 subjects; variants (and their frequencies) included 28M/V (1%), 30A/K/S/T/V (10%), 31L/M (1%), E92A (1%), and Y93H (8.3%). A heterozygous JFH1/3a-NS5A replicon was constructed using a shared sequence and its sensitivity to NS5A receptor antagonists was evaluated. For JFH1/3a-NS5A, DCV showed higher potency (EC50 = 0.52 nM) than GS-5885 (EC50 = 141 nM). DCV exhibited enhanced sensitivity to JFH1/3a-NS5A-M28V (EC50 = 0.006 nM), A30V (EC50 = 0.012 nM), and E92A (EC50 = 0.004 nM), while the NS5A-A30K and -Y93H variants showed decreased sensitivity to DCV (EC50 values of 23 nM and 1120 nM, respectively) and GS-5885 (EC50 values of 1770 nM and 4300 nM, respectively). Conclusion: Amino acid substitutions that confer resistance to NS5A blocking inhibitors have been found in untreated HCV GT3-infected patients. The clinical efficacy of these NS5A blocking inhibitors may depend on the type of inhibitor used in combination with other antiviral drugs. [2]

C53H60F2N8O12

1039.09

1038.43

1502654-87-6

Ledipasvir;1256388-51-8;Ledipasvir (acetone);1441674-54-9;Ledipasvir-d6;2050041-12-6;Ledipasvir hydrochloride;2128695-48-5;Ledipasvir (diacetone);1502655-48-2

78357794

Off-white to yellow solid powder

7.142

8

16

15

75

1950

8

CC(C)[C@@H](C(=O)N1CC2(CC2)C[C@H]1C3=NC=C(N3)C4=CC5=C(C=C4)C6=C(C5(F)F)C=C(C=C6)C7=CC8=C(C=C7)N=C(N8)[C@@H]9[C@H]1CC[C@H](C1)N9C(=O)[C@H](C(C)C)NC(=O)OC)NC(=O)OC.[C@H]([C@@H](C(=O)O)O)(C(=O)O)O

ZQVLPYMRXLPMDX-KEAIDYLOSA-N

InChI=1S/C49H54F2N8O6.C4H6O6/c1-24(2)39(56-46(62)64-5)44(60)58-23-48(15-16-48)21-38(58)42-52-22-37(55-42)28-9-13-32-31-12-8-26(18-33(31)49(50,51)34(32)19-28)27-10-14-35-36(20-27)54-43(53-35)41-29-7-11-30(17-29)59(41)45(61)40(25(3)4)57-47(63)65-6;5-1(3(7)8)2(6)4(9)10/h8-10,12-14,18-20,22,24-25,29-30,38-41H,7,11,15-17,21,23H2,1-6H3,(H,52,55)(H,53,54)(H,56,62)(H,57,63);1-2,5-6H,(H,7,8)(H,9,10)/t29-,30+,38-,39-,40-,41-;1-,2-/m00/s1

Methyl N-[(2S)-1-[(6S)-6-[5-[9,9-Difluoro-7-[2-[(1S,2S,4R)-3-[(2S)-2-(methoxycarbonylamino)-3-methylbutanoyl]-3-azabicyclo[2.2.1]heptan-2-yl]-3H-benzimidazol-5-yl]fluoren-2-yl]-1H-imidazol-2-yl]-5-azaspiro[2.4]heptan-5-yl]-3-methyl-1-oxobutan-2-yl]carbamate tartrate

GS-5885 tartrate, GS5885 tartrate; GS 5885; Ledipasvir D-tartrate; Ledipasvir (D-tartrate); 1502654-87-6; RT680T6HCQ; 1499193-68-8; UNII-RT680T6HCQ; (2S,3S)-2,3-dihydroxybutanedioic acid;methyl N-[(2S)-1-[(6S)-6-[5-[9,9-difluoro-7-[2-[(1R,3S,4S)-2-[(2S)-2-(methoxycarbonylamino)-3-methylbutanoyl]-2-azabicyclo[2.2.1]heptan-3-yl]-3H-benzimidazol-5-yl]fluoren-2-yl]-1H-imidazol-2-yl]-5-azaspiro[2.4]heptan-5-yl]-3-methyl-1-oxobutan-2-yl]carbamate; trade name: Harvoni;

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 : ~25 mg/mL (~24.06 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (2.41 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 (2.41 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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Solubility in Formulation 3: 10% DMSO+90% Corn Oil: ≥ 2.5 mg/mL (2.41 mM)

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