L-type calcium channel (Ca2+ channel blocker)[1]

In vitro activity: Manidipine, a Ca(2+)-channel blocker, at concentrations that lower elevated blood pressure, modulates the transcription rates of cytokine genes in the mesangial cells of humans that had been stimulated with platelet-derived growth factor BB isomer. Manidipine inhibits the ET-1-induced [Ca2+]i increase by reducing both the transient and sustained Ca2+ increments in A7r5 cells and glomerular mesangial cells (MCs). Manidipine (10(-5) mol/L) potentiates ET-1-induced c-fos and c-jun expression in A7r5 cells. Manidipine is a potent inhibitor for ET-1-induced [Ca2+]i signaling and that Manidipine has multiple effects on ET-1-induced signaling, including potentiating the immediate-early gene response.

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Manidipine inhibited PDGF-BB-induced DNA synthesis in human mesangial cells with an IC50 of 4 × 10^{-8} M after 24 hr of incubation.[1]
Manidipine inhibited PDGF-BB-induced cell proliferation with an IC50 of 6 × 10^{-8} M after 3 days of culture.[1]
At concentrations from 10^{-9} to 10^{-6} M, Manidipine (10^{-8} M) inhibited PDGF-BB-induced IL-1β mRNA transcription to basal level, inhibited GM-CSF mRNA transcription by approximately 50%, and enhanced IL-6 mRNA transcription by 1.5-fold relative to PDGF-BB alone.[1]
Manidipine (10^{-8} M) abolished PDGF-BB-induced c-fos and HMG-CoA reductase mRNA transcription, and partially reduced c-jun mRNA transcription by about 50%, as shown by Northern blot analysis.[1]
Nuclear runoff assays confirmed that Manidipine affects mRNA levels at the level of transcription rather than by altering degradation; the increase in IL-6 steady-state mRNA was abolished by actinomycin D (5 μg/ml).[1]
The viability of mesangial cells treated with Manidipine was approximately 95% after 24 hr of culture; cells washed and re-exposed to PDGF-BB responded normally in DNA synthesis and cell growth, and morphology was unaltered by phase-contrast microscopy.[1]

Manidipine reduces systolic blood pressure with a slight sympathetic reflex increase in heart rate, and increases plasma nitrite/nitrates in perfused rat heart from ischemia-reperfusion damage. Manidipine combined with Simvastatin reduces creatine kinase, lactate dehydrogenase and tumor necrosis factor-alpha, and enhancement of 6-keto-PGF(1alpha) during reperfusion. Manidipine hydrochloride prevents isoproterenol-induced left ventricular hypertrophy and expression of mRNA of ANP, collagen types I and type III, and fibronectin in rats with isoproterenol-induced cardiac hypertrophy. Manidipine HCl increases renal blood flow (RBF) by dilating the afferent arterioles and improves glomerular hypertension by dilating the efferent arterioles in hypertensive rats.

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Human mesangial cells (two primary lines H43, H44) were cultivated in RPMI 1640 medium supplemented with 10% fetal calf serum, 1% vitamin mix, 8 mM L-glutamine, and insulin/transferrin/selenite additive. Two days prior to experiments, 1×10^6 cells were seeded into 750 ml flasks; 4 hr later the medium was exchanged with RPMI 1640 containing 0.1% fetal calf serum (low serum condition) to render cells quiescent, and the low serum medium was changed daily until stimulation.[1]
For mitogenicity measurement, 1×10^5 quiescent cells were seeded into 25 ml flasks in low serum medium. On the following day, cells were preincubated with Manidipine (1×10^{-8} M) for 3 hr, then stimulated with PDGF-BB (10 ng/ml) or kept in low serum medium alone. Medium was replaced daily, and cells were counted on days 1, 3, and 5. The mitogenic effect was also measured by ³H-thymidine incorporation into DNA. Experiments were performed in triplicate using both cell lines.[1]
Total RNA was extracted according to the method of Chomczynski and Sacchi, quantitated by spectrophotometry (260/280 nm). For Northern blot analysis, equal amounts of RNA were denatured at 95°C for 5 min in sample buffer (50% formamide, 10% glycine, 1% bromophenol blue), then size-fractionated by electrophoresis through 1% agarose gel containing 0.02 M morpholinosulfonate, 2% formaldehyde, and 0.2 μg/ml ethidium bromide, followed by transfer and hybridization.[1]
Nuclear runoff assays were performed as described by Nevins. Nuclei from 1×10^8 mesangial cells were isolated, and RNA chains were elongated with creatine kinase (5 mg/ml) in the presence of 500 mCi of ³²P-UTP (3000 mCi/mmol) and 10 mM each of ATP, CTP, and GTP. Labeled RNA was extracted by the guanidinium isothiocyanate method and analyzed by dot blot. Quantitation of de novo mRNA synthesis was assessed using autoradiography.[1]

Rats

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Absorption, Distribution and Excretion
The median time to peak concentration (Tmax) is 1.5 hours. Co-administration with food increases Cmax by 1.3–1.6 times, but does not change Tmax. Manidipine does not accumulate significantly after multiple doses. Manidipine is primarily eliminated through extensive metabolism. 63% of the drug is excreted in feces as metabolites, and 31% in urine. Metabolism/Metabolites Manidipine is primarily metabolized by CYP enzymes to pyridine derivatives and diphenylmethane derivatives, accounting for 4–7% and 22–24% of the urinary excretion, respectively. Biological Half-Life Elimination half-life has been observed to be dose-dependent. Half-lives of 3.94, 5.02, and 7.95 hours were observed at doses of 5, 10, and 20 mg, respectively.

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Protein Binding
Manidipine binds to human plasma proteins at a rate of 99%.

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Manidipine-treated mesangial cells showed approximately 95% viability after 24 hr of culture. Cells washed and re-exposed to PDGF-BB responded normally in terms of DNA synthesis and cell growth. Phase-contrast microscopy revealed no alteration in cell morphology.[1]

Proc Natl Acad Sci U S A.1992;89(9):4071-5;Cancer Res.1992;52(5):1369-71.

Manidipine is a diarylmethane compound. Manidipine (INN) is a calcium channel blocker (dihydropyridine) used clinically as an antihypertensive drug. It is vascular selective and has no cardiac effects at clinically relevant doses. Indications: For the treatment of hypertension. Mechanism of Action: Vascular smooth muscle contraction is caused by stimulation of Gq-coupled receptors, which lead to the release of calcium ions from the sarcoplasmic reticulum. Subsequently, voltage-dependent calcium channels open, allowing calcium ions to flow into the cells, ultimately resulting in vasoconstriction. Manidipine slowly binds to and dissociates from L- and T-type voltage-dependent calcium channels on smooth muscle cells, blocking the entry of extracellular calcium ions into the cells, thereby preventing vasoconstriction. This leads to vasodilation and lowers blood pressure. Manidipine can cause renal vasodilation and increased sodium excretion. This may exert its antihypertensive effect by reducing blood volume. Manidipine is vascular selective and has no significant effects on the heart or central nervous system at clinically relevant doses.

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Pharmacodynamics
Manidipine can cause vasodilation, thereby lowering blood pressure.

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Manidipine at nanomolar concentrations modulates gene transcription involved in proinflammatory changes of mesangial cells, suggesting anti-inflammatory and protective effects in glomerular diseases. Its effects are independent of Ca2+ influx, as neither BayK 8644 (Ca2+ channel opener), TMB-8 (intracellular Ca2+ release blocker), Ca2+-free medium, nor the Ca2+ ionophore A23187 altered the modulation of PDGF-dependent gene transcription by Manidipine.[1]
The drug's action involves protein kinase C (PKC), because (i) PDGF stimulates PKC activation; (ii) PDGF induces c-fos and HMG-CoA reductase, which are known to be stimulated by PKC; (iii) Manidipine blocks these inductions; and (iv) PDGF-dependent PKC activation can be inhibited by Ca2+-channel blockers. Manidipine (10^{-8} M) abolished PDGF-induced c-fos and HMG-CoA reductase transcription, similar to the PKC inhibitor polymyxin B.[1]
Manidipine may affect the activity of PKC by intercalating into biomembranes, thereby altering hydrophobic/hydrophilic interactions, and consequently influence AP-1 (Fos/Jun) transcription factor activity. The ability to alter c-jun/AP-1 activation provides a modulatory influence on gene activation.[1]
Compared to other Ca2+-channel blockers (verapamil, diltiazem, amlodipine) at 10^{-8} M, Manidipine and verapamil enhanced IL-6 transcription while diltiazem and amlodipine were ineffective; all four inhibited IL-1β transcription to basal level and inhibited GM-CSF transcription by ~50%.[1]
Manidipine at therapeutic (nanomolar) concentrations lowers elevated blood pressure and modulates transcription of genes involved in the inflammatory response of mesangial cells, potentially protecting against renal injury and atherosclerosis.[1]

C35H38N4O6

610.7

610.279

89226-50-6

Manidipine dihydrochloride;89226-75-5;Manidipine-d4;1189656-59-4

4008

Light yellow to yellow solid powder

1.2±0.1 g/cm3

722.0±60.0 °C at 760 mmHg

125-128ºC

390.4±32.9 °C

0.0±2.3 mmHg at 25°C

1.601

5.46

1

9

11

45

1090

0

O=C(C1C(C2C=C([N+](=O)[O-])C=CC=2)C(C(OCCN2CCN(C(C3C=CC=CC=3)C3C=CC=CC=3)CC2)=O)=C(C)NC=1C)OC

JINNGBXKBDUGQT-UHFFFAOYSA-N

InChI=1S/C35H38N4O6.2ClH/c1-24-30(34(40)44-3)32(28-15-10-16-29(23-28)39(42)43)31(25(2)36-24)35(41)45-22-21-37-17-19-38(20-18-37)33(26-11-6-4-7-12-26)27-13-8-5-9-14-27;;/h4-16,23,32-33,36H,17-22H2,1-3H3;2*1H

3-(2-(4-benzhydrylpiperazin-1-yl)ethyl) 5-methyl 2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate dihydrochloride

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)

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.)