SCY-635, a 3,4-disubstituted nonimmunosuppressive CsA analog that exhibits potent suppression of HCV RNA replication in vitro. SCY-635 is a reversible, nanomolar inhibitor of the PPIase activity expressed by CyPA. SCY-635 is orally bioavailable in multiple species and distributes extensively to hepatocytes. Metabolic studies indicate that the administration of SCY-635 is unlikely to result in adverse pharmacological interactions. Two-drug synergy studies indicate that SCY-635 exhibits additive to synergistic antiviral activity when it is tested in vitro with alpha interferon 2b (IFNα-2b) or ribavirin without increasing cell cytotoxicity. These results are consistent with the recent observation of the potent clinical antiviral activity of SCY-635 (13) and suggest that further clinical studies assessing the safety and antiviral activity of SCY-635 in combination with interferon and ribavirin are warranted.

Culture medium was supplemented either with 5% fetal bovine serum (FBS) (0% human serum) or with human serum to achieve final concentrations of 10%, 20%, and 40%. Anti-HCV activity (expressed as the 50% effective concentration [EC50]) was assessed by using the luciferase end point following 72 h of incubation with SCY-635 at concentrations ranging from 0.05 to 5 μM.[1]
CsA or SCY-635 was incubated in a 1:1 molar ratio with CyPA in the presence of calcineurin (1.32 nM) and calmodulin (50 nM) at 22°C for 30 min. [1]
CsA, SCY-635, or a control compound (warfarin, imipramine, or carbamezapine) was added to pooled fresh human plasma , and the mixture was incubated at 37°C for 30 min before centrifugation. The amounts of unbound (free) and bound compound in the ultrafiltrate and retentate, respectively, were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Briefly, chromatographic separation was achieved by gradient elution from a Luna C8 3-μm-particle-size HPLC column (50 by 2 mm). The starting gradient conditions were 80:20 water-methanol (5 mM ammonium formate and 0.1% formic acid) for 15 s, followed by a linear gradient to 5:95 water-methanol over 1.75 min and a hold for 1 min. SCY-635 eluted at 2.3 min. [1]
CsA and SCY-635 (1 μM) were incubated with pooled liver microsomes (1.05 mg protein/ml) for 0, 5, 15, and 30 min at 37°C in an oxygen-enriched environment in the presence of NADPH.[1]
CsA and SCY-635 were added to membrane preparations containing recombinant human CYP enzymes at concentrations ranging from 1 to 100 μM, together with luminogenic substrates specific for each enzyme. [1]
SCY-635 was tested at nine concentrations with rIFNα-2b prepared at five concentrations. SCY-635 was serially diluted twofold with concentrations ranging from 0.008 to 2.0 μM. rIFNα-2b was prepared as half-log10 dilutions, producing concentrations of 0.005 to 0.5 IU/ml. SCY-635 was tested at eight concentrations with ribavirin tested at five concentrations. SCY-635 was prepared by the use of twofold dilutions, with the resulting concentrations ranging from 3.91 to 500 nM. [1]
The hepatocytes were seeded in the bottom of a 24-well Costar plate at a density of 2 × 105 cells/well. A Transwell insert plate was carefully added and the NPCs were seeded in each insert at a density of 1 × 105 cells per well. SCY-635 was added at a final concentration of 500 or 3,000 ng/ml. The plates were incubated for 1 h at 37°C in a humidified incubator with 5% CO2. The cells were collected at the end of the incubation. The protein content was measured as described by Lowry . The concentration of SCY-635 in the hepatocytes and nonparenchymal cells was determined by LC-MS/MS. The SCY-635 concentration was expressed as the ng of SCY-635 per μg of protein.[1]

The biological distribution of SCY-635 in male Sprague-Dawley rats was assessed following the intravenous injection of 10 mg/kg of body weight and following oral gavage with 10 and 30 mg/kg. [1]

Antimicrob Agents Chemother.2010 Feb;54(2):660-72;Antimicrob Agents Chemother.2012 Jul;56(7):3888-97;J Hepatol.2012 Jul;57(1):47-54.

SCY-635 has been investigated for the treatment of Chronic Hepatitis C and Hepatitis C Infection.

C66H120N12O13S

1321.8

1320.88

210759-10-7

6479267

Typically exists as solid at room temperature

3.885

6

15

19

92

2530

13

C/C=C/C[C@H]([C@@H](O)[C@H]1C(N[C@H](C(N([C@H](SCCN(C)C)C(N([C@H](C(N[C@H](C(N([C@H](C(N[C@H](C(N[C@@H](C(N([C@H](C(N([C@H](C(N([C@H](C(N1C)=O)C(C)C)C)=O)CC(C)C)C)=O)CC(C)C)C)=O)C)=O)C)=O)CC(C)C)C)=O)C(C)C)=O)CC(C)(O)C)C)=O)C)=O)CC)=O)C

AQHMBDAHQGYLIU-XNFHFXFQSA-N

InChI=1S/C66H120N12O13S/c1-27-29-30-42(13)53(79)52-57(83)69-45(28-2)59(85)78(26)65(92-32-31-71(18)19)64(90)75(23)49(36-66(16,17)91)56(82)70-50(40(9)10)62(88)72(20)46(33-37(3)4)55(81)67-43(14)54(80)68-44(15)58(84)73(21)47(34-38(5)6)60(86)74(22)48(35-39(7)8)61(87)76(24)51(41(11)12)63(89)77(52)25/h27,29,37-53,65,79,91H,28,30-36H2,1-26H3,(H,67,81)(H,68,80)(H,69,83)(H,70,82)/b29-27+/t42-,43+,44-,45+,46+,47+,48+,49+,50+,51+,52+,53-,65-/m1/s1

(3S,6S,9S,12R,15S,18S,21S,24S,27R,30S,33S)-27-[2-(dimethylamino)ethylsulfanyl]-30-ethyl-33-[(E,1R,2R)-1-hydroxy-2-methylhex-4-enyl]-24-(2-hydroxy-2-methylpropyl)-1,4,7,10,12,15,19,25,28-nonamethyl-6,9,18-tris(2-methylpropyl)-3,21-di(propan-2-yl)-1,4,7,10,13,16,19,22,25,28,31-undecazacyclotritriacontane-2,5,8,11,14,17,20,23,26,29,32-undecone

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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