| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
Adenylate cyclase (catalytic unit) – directly activates the catalytic subunit of adenylate cyclase and increases intracellular cAMP [1].
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| ln Vitro |
Lymphocytes isolated from mesenteric lymph nodes of NKH477‑treated allograft recipients (3 mg/kg/day for 5 days) showed inhibited spontaneous proliferation when cultured alone, compared to untreated recipients. Re‑stimulation with donor antigens (x‑irradiated donor splenocytes) for 72 hours also resulted in suppressed proliferative response in the NKH477‑treated group. These ex vivo results reflect an in vivo antiproliferative effect of NKH477 on lymphocytes [1].
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| ln Vivo |
In a dose-dependent manner, NKH477 (Colforsin dapropate hydrochloride) (oral; 1-3 mg/kg/day; days 3 and 5) improves lung allograft survival [1].
Oral administration of NKH477 to rat lung allograft recipients (Brown‑Norway to Lewis) significantly prolonged graft survival in a dose‑dependent manner. Mean survival times (MST): untreated = 4.3 days; NKH477 1 mg/kg/day = 5.7 days; 2 mg/kg/day = 8.2 days; 3 mg/kg/day = 9.5 days (p < 0.01 vs untreated) [1]. Histopathological rejection grades were significantly reduced by NKH477 (3 mg/kg/day) at both post‑operative day 3 and day 5 (grade 1.4±0.5 and 2.4±0.5, respectively) compared to untreated allografts (2.4±0.5 and 3.6±0.5) (p < 0.05) [1]. In lung allografts, NKH477 suppressed the upregulation of IFN‑γ and IL‑10 measured by ELISA. At day 3 and day 5, IFN‑γ and IL‑10 levels were markedly lower in NKH477‑treated grafts than in untreated allografts (p < 0.05). Expression of IL‑2 and IL‑4 in grafts was not altered by NKH477 [1]. In recipient spleens (immunohistochemistry), NKH477 suppressed alloresponsive expression of IL‑2 and IL‑10 at day 5 (p < 0.01 and p < 0.05, respectively). Splenic IFN‑γ expression was also abrogated, while IL‑4 expression was not significantly changed [1]. |
| Cell Assay |
Mixed lymphocyte reaction (MLR) – Mononuclear cells from naive donor rats (Brown‑Norway) were obtained by density gradient centrifugation of splenocytes and x‑irradiated (3000 rad). Mesenteric lymph node cells were isolated from recipient rats (Lewis) that were either untreated or treated daily with NKH477 (3 mg/kg/day) for 5 days after lung allotransplantation. Cells (2.5×10^5/well) were cultured alone or stimulated with x‑irradiated donor splenocytes (2.5×10^5/well) in 200 μL medium in 96‑well round‑bottom plates for 72 hours. Cultures were pulsed with 1 μCi/well [³H]thymidine for the last 6 hours, then harvested onto glass‑fiber filters. Beta emissions were measured as cpm using a beta‑scintillation counter. Assays were performed in triplicate [1].
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| Animal Protocol |
Animal/Disease Models: Specific pathogen-free inbred male Lewis rat (LEW), weighing 250 to 280 g [1]
Doses: 1, 2 or 3 mg/kg/day Doses: Oral; daily; day 3 and Day 5 Experimental Results: Lung allograft survival was dose-dependently prolonged. Orthotopic left lung transplantation from Brown‑Norway donors to Lewis recipients. Recipients were treated orally once daily from day 0 to day 10 post‑transplantation or until rejection. Drug preparation: NKH477 powder dissolved in saline and adjusted to pH 4 with 0.01 N HCl, prepared daily and stored at 4°C. Doses used: 1, 2, or 3 mg/kg body weight per day. Control groups: untreated allografts and isografts (Lewis to Lewis) [1]. Graft function monitored daily by chest roentgenography using a ventilation score (6 = normal to 0 = opaque); rejection defined as score 1 or 0 [1]. Histopathology: recipients sacrificed on day 3 or day 5; transplanted lungs flushed with buffered formalin, embedded in paraffin, sections stained with hematoxylin and eosin, rejection graded per ISHLT classification by blinded pathologists [1]. Cytokine analysis: lung grafts homogenized in saline, centrifuged, supernatant used for ELISA (IFN‑γ, IL‑2, IL‑4, IL‑10). Spleens sliced, embedded in OCT, frozen, cryosections (4‑6 μm) fixed in acetone, stained with primary antibodies against rat IL‑2, IFN‑γ, IL‑4, and IL‑10, then biotinylated secondary antibodies and avidin‑conjugated alkaline phosphatase; staining scored 0‑3 by blinded observers [1]. Ex vivo MLR as described in Cell Assay section [1]. |
| References | |
| Additional Infomation |
Cefosin darope ester hydrochloride is the hydrochloride salt of cefosin darope ester. It is a drug used to treat acute heart failure. It directly stimulates adenylate cyclase, producing a positive inotropic effect and vasodilatory effect, accompanied by an increase in intracellular cAMP levels. It has the effects of adenylate cyclase agonist, antihypertensive drug, cardiotonic drug, and vasodilator. It contains cefosin darope ester (1+). It potently activates the adenylate cyclase system and cyclic adenosine monophosphate (cAMP) biosynthesis. Extracted from Coleus forskohlii. It has antihypertensive, positive inotropic, platelet aggregation inhibitory, and smooth muscle relaxant effects; it can also lower intraocular pressure and promote the release of pituitary hormones.
NKH477 is a water‑soluble forskolin derivative ((+)-(3R,4aR,5S,6S,6aS,10S,10aR,10bS)-5-acetoxy-6-(3-dimethylamino-propionyloxy)-dodecahydro-10,10b-dihydroxy-3,4a,7,7,10a-pentamethyl-3-vinyl-1H-naphtho-[2,1-b]pyran-1-one monohydrochloride) originally introduced as a positive inotropic agent for heart failure. Like forskolin, it directly activates adenylate cyclase and increases intracellular cAMP. Its hydrophobic nature and putative non‑specific activities had hindered in vivo use, but NKH477 overcomes solubility issues [1]. The drug has been reported to prolong survival of murine cardiac allografts prior to this study. This study demonstrates for the first time that oral NKH477 exerts antiproliferative effects on lymphocytes in vivo with an altered cytokine profile (inhibition of alloresponsive IL‑2 and IFN‑γ), leading to reduced acute rejection and prolonged survival of rat lung allografts [1]. |
| Molecular Formula |
C28H45CLO8
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|---|---|
| Molecular Weight |
545.11
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| Exact Mass |
545.275
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| CAS # |
138605-00-2
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| PubChem CID |
444028
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| Appearance |
White to off-white solid powder
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| Boiling Point |
571.4ºC at 760mmHg
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| Flash Point |
176ºC
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| Vapour Pressure |
5.29E-16mmHg at 25°C
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| LogP |
2.824
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
37
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| Complexity |
933
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| Defined Atom Stereocenter Count |
8
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| SMILES |
C=C[C@@]1(C)CC(=O)[C@@]2([C@@]3(C)[C@H](CCC(C)(C)[C@@H]3[C@@H]([C@@H]([C@@]2(C)O1)OC(=O)C)OC(=O)CCN(C)C)O)O.Cl
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| InChi Key |
LNEICJSEZPSKSJ-RPBMHHKSSA-N
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| InChi Code |
InChI=1S/C28H44O8.ClH/c1-10-25(7)15-19(31)28(33)26(8)18(30)13-14-24(5,6)22(26)21(35-20(32)12-11-16(2)3)23(34-17(4)29)27(28,9)36-25/h10,16,18,21-23,30,33H,1,11-15H2,2-9H31H/t18-,21-,22-,23-,25-,26-,27+,28-/m0./s1
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| Chemical Name |
(3R,4aR,5S,6S,6aS,10S,10aR,10bS)-5-(acetyloxy)-3-ethenyl-10,10b-dihydroxy-3,4a,7,7,10a-pentamethyl-1-oxo-dodecahydro-1H-naphtho[2,1-b]pyran-6-yl 4-methylpentanoate hydrochloride
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| Synonyms |
NKH 477 NKH-477 NKH477
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. (3). This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~228.90 mM)
H2O : ~20 mg/mL (~36.62 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.81 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. Solubility in Formulation 2: ≥ 2.08 mg/mL (3.81 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (3.81 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8345 mL | 9.1725 mL | 18.3449 mL | |
| 5 mM | 0.3669 mL | 1.8345 mL | 3.6690 mL | |
| 10 mM | 0.1834 mL | 0.9172 mL | 1.8345 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT00864578 | WITHDRAWN | Intraocular Pressure Primary Open Angle Glaucoma |
Sooft Italia | 2009-04 | ||
| NCT02143349 | UNKNOWN STATUS | Dietary Supplement: Coleus forskohlii extract
Other: Placebo |
Metabolic Syndrome Obesity |
Olive Lifesciences Pvt Ltd | 2014-04 | Phase 3 |
| NCT03652090 | COMPLETED | Procedure: cell sampling | Cystic Fibrosis | Institut National de la Santé Et de la Recherche Médicale, France | 2010-09-01 | |
| NCT04254705 | WITHDRAWN | Procedure: Rectal Biopsy | Cystic Fibrosis | Universitaire Ziekenhuizen KU Leuven | 2020-03-01 | Not Applicable |
| NCT01254006 | COMPLETED | Drug: forskolin, rutin and vitamins B1 and B2 | Glaucoma | University of Roma La Sapienza | Not Applicable |
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