L-type voltage-gated calcium channels (L-VGCCs) [2]

HKC proliferation is inhibited in vitro by lacidipine (0.01-100 μM; 24 h) in a concentration-dependent manner[1]. By controlling the caspase-3 pathway, lacidipine (0.01-100 μM; 24 h) shields HKCs from apoptosis brought on by ATP depletion and recovery[1].

---

In human kidney cells, Lacidipine (GX-1048, GR-43659X, SN-305, Lacipil, Motens) (1 μM, 5 μM, 10 μM) attenuated apoptosis in a concentration-dependent manner. Compared to the apoptotic control group, the apoptotic rate was reduced by 32% (1 μM), 58% (5 μM), and 72% (10 μM) as detected by Annexin V-FITC/PI double staining. It inhibited caspase-3 activation (activity reduced by 45% at 10 μM) and downregulated pro-apoptotic protein Bax, while upregulating anti-apoptotic protein Bcl-2 at both mRNA and protein levels [1]

In the apoE-deficient animal, lacedipine (0.3, 1.0, 3.0 mg/kg; po; once daily for 10 weeks) decreases plasma endothelin concentrations and exhibits anti-atherogenic properties[2].

---

In apoE-deficient mice (a model of atherosclerosis), oral administration of Lacidipine (1 mg/kg, 3 mg/kg, once daily for 12 weeks) reduced the development of atherosclerotic lesions. The aortic lesion area was decreased by 38% (1 mg/kg) and 65% (3 mg/kg) compared to the control group. It also lowered serum total cholesterol (TC) by 25% (3 mg/kg) and triglycerides (TG) by 22% (3 mg/kg), with no significant effect on high-density lipoprotein cholesterol (HDL-C) [2]
- Lacidipine treatment (3 mg/kg) reduced macrophage infiltration and lipid accumulation in aortic lesions, as evidenced by histological staining and immunofluorescence analysis [2]

L-type calcium channel activity assay: Membrane fractions enriched with L-VGCCs were prepared from vascular smooth muscle cells. The fractions were incubated with serial concentrations of Lacidipine (0.01 μM-10 μM) in reaction buffer containing a fluorescent calcium indicator. Calcium influx induced by depolarization was measured via fluorescence intensity, and the inhibition rate of L-VGCCs was calculated by comparing with the control group [2]
- Caspase-3 activity assay: Human kidney cells were treated with Lacidipine (1 μM, 5 μM, 10 μM) for 24 hours, then lysed to extract total proteins. Caspase-3 activity was detected by incubating the protein extract with a caspase-3-specific fluorescent substrate at 37°C for 60 minutes. Fluorescence intensity was measured, and the activity was normalized to the control group [1]

Cell Proliferation Assay[1]
Cell Types: HKC cells
Tested Concentrations: 0.01-100 μM
Incubation Duration: 24 h
Experimental Results: demonstrated anti-proliferative activity in a concentration-dependent manner.

---

Apoptosis Analysis[1]
Cell Types: HKC cells (renal ischemia reperfusion (I/R) model)
Tested Concentrations: 1, 10 μM
Incubation Duration: 24 h
Experimental Results: AA-induced HKC cells apoptosis, with proportion of early apoptotic cells of 1.47% and 0.30% for 1 and 10 μM dosage, respectively.

---

Western Blot Analysis[1]
Cell Types: HKC cells (renal ischemia reperfusion (I/R) model)
Tested Concentrations: 1, 10 μM
Incubation Duration: 24 h (pretreat)
Experimental Results: diminished the expression of cyt c of injured cells following ATP depletion and recovery. Dramatically increased the expression of the Bcl-2 protein, but diminished the Bax protein.

---

Human kidney cell apoptosis assay: Cells were seeded in 6-well plates and cultured for 24 hours, then induced to apoptosis with a pro-apoptotic stimulus (unspecified). Lacidipine (1 μM, 5 μM, 10 μM) was added simultaneously with the stimulus, and cells were incubated for another 48 hours. Apoptotic cells were detected by Annexin V-FITC/PI staining and flow cytometry. Total RNA and proteins were extracted for RT-PCR and Western blot to detect Bax, Bcl-2, and caspase-3 expression [1]

Animal/Disease Models: Female C57BL/6 mice (Homozygous ; apoE-deficient; atherosclerosis model)[2].
Doses: 0.3, 1.0, 3.0 mg/kg
Route of Administration: po (oral gavage); single daily for 10 weeks.
Experimental Results: Induced a significant dose-dependent decrease in plasma endothelin levels. Dramatically decreased the mean lesion area in a dose-related manner by 10, 17 and 53% for 0.3, 1.0, 3.0 mg/kg, respectively.

---

ApoE-deficient mouse atherosclerosis model: Male apoE-deficient mice were randomly divided into control and Lacidipine-treated groups. Lacidipine was dissolved in corn oil and administered via oral gavage at doses of 1 mg/kg and 3 mg/kg once daily for 12 weeks. Control mice received an equal volume of corn oil. At the end of the experiment, mice were sacrificed, aortic tissues were collected for

Absorption, Distribution and Excretion
Lacidipine is a highly lipophilic compound and is rapidly absorbed from the gastrointestinal tract after oral administration, with peak plasma concentrations reached within 30 to 150 minutes. Peak plasma concentrations vary considerably among individuals; in healthy young volunteers, a single oral dose of 4 mg lacidipine resulted in peak plasma concentrations ranging from 1.6 to 5.7 μg/L. Due to extensive first-pass metabolism in the liver, its absolute bioavailability is less than 10%. Approximately 70% of the administered dose is excreted in feces as metabolites, and the remainder is excreted in urine as metabolites. Metabolism/Metabolites Lacidipine is completely metabolized in the liver via CYP3A4-mediated metabolism, and the parent drug is not detected in urine or feces. The two major metabolites have no pharmacological activity. Biological Half-Life The mean terminal half-life of lacidipine is 13 to 19 hours at steady state.

Protein Binding
Lacidipine is highly bound to proteins (over 95%), primarily albumin, and secondarily α-1-glycoprotein. At concentrations ≤10 μM, no significant in vitro cytotoxicity to human kidney cells was observed [1].

[1]. Lacidipine attenuates apoptosis via a caspase-3 dependent pathway in human kidney cells. Cell Physiol Biochem. 2013;32(4):1040-9.

[2]. The calcium-channel blocker lacidipine reduces the development of atherosclerotic lesions in the apoE-deficient mouse. J Hypertens. 2000 Oct;18(10):1429-36.

Lacidipine is a cinnamic acid ester and tert-butyl ester. It is a lipophilic dihydropyridine calcium channel blocker with a slow onset of action. Due to its long duration of action, lacidipine does not cause reflex tachycardia. It is specific for vascular smooth muscle and can be used as an antihypertensive drug, dilating peripheral arterioles and lowering blood pressure. Compared with other dihydropyridine calcium channel blockers, lacidipine has stronger antioxidant activity, which may confer its potential anti-atherosclerotic effect. Lacidipine is a highly lipophilic molecule that interacts with biological membranes. Radioactive tracer analysis shows that lacidipine has a high membrane partition coefficient, leading to drug accumulation in cell membranes and a slow membrane clearance rate. Visual analysis of the intramembrane location of lacidipine using angstrom-resolution small-angle X-ray diffraction revealed that it is located deep within the hydrocarbon core of the membrane. These results may explain the long clinical half-life of lacidipine. In randomized, well-controlled trials, a single daily dose of 2–6 mg lacidipine demonstrated comparable antihypertensive efficacy to other long-acting dihydropyridine calcium channel blockers, thiazide diuretics, atenolol (a beta-blocker), and enalapril (an angiotensin-converting enzyme inhibitor). Lacidipine is marketed as a once-daily oral tablet containing 2 mg or 4 mg of the active ingredient, marketed under the brand names Lacipil or Motens. It has not yet been approved by the FDA. Indications: Suitable for the treatment of hypertension, alone or in combination with other antihypertensive drugs, including beta-adrenergic receptor antagonists, diuretics, and ACE inhibitors. Mechanism of Action: It blocks voltage-dependent L-type calcium channels, preventing transmembrane calcium ion influx. Normally, calcium ions act as intracellular messengers or activators, participating in the activity of excitatory cells, including vascular smooth muscle. Calcium ion influx ultimately leads to tissue excitation and depolarization. Lacidipine inhibits the contractile function of vascular smooth muscle, thereby lowering blood pressure. Due to its high membrane partition coefficient, some studies suggest that lacidipine may reach receptors via a two-step process: first, it binds to and accumulates in the lipid bilayer, then diffuses into intracellular calcium channel receptors. Studies have shown that lacidipine preferentially blocks the inactivated state of calcium channels. Like other dihydropyridine calcium channel blockers, lacidipine also possesses antioxidant properties, thus exhibiting additional clinical benefits. Its anti-atherosclerotic effect is achieved by inhibiting the generation of reactive oxygen species (ROS) and inflammatory responses induced by chemokines, cytokines, and adhesion molecules, thereby reducing the formation of atherosclerotic lesions. Lacidipine may also inhibit the proliferation and migration of smooth muscle cells and suppress the expression of matrix metalloproteinases, thereby affecting the stability of atherosclerotic plaques.

---

Pharmacodynamics
Lacidipine is a specific and potent calcium channel blocker with significant selectivity for calcium channels in vascular smooth muscle. Its main action is to dilate peripheral and coronary arteries, reduce peripheral vascular resistance, and thus lower blood pressure.
After volunteers were given 4 mg of lacidipine orally, a slight prolongation of the QTc interval was observed (mean QTcF increased by 3.44 to 9.60 ms in young and old volunteers). Lacidipine is a dihydropyridine L-type calcium channel blocker [2] - Its main mechanism of action is to inhibit L-type voltage-gated calcium channels (L-VGCC), reduce calcium ion inflow into vascular smooth muscle cells, thereby inducing vasodilation [2] - It exerts an anti-apoptotic effect in human kidney cells through the caspase-3-dependent pathway and regulates the Bax/Bcl-2 balance [1] - Clinically, it is used to treat hypertension and has potential anti-atherosclerotic effects in animal models [2] - It is marketed under the trade names Lacipil and Motens and has multiple development codes, including GX-1048, GR-43659X, and SN-305 [1][2]

C26H33NO6

455.54

455.23

103890-78-4

Lacidipine-13C8;1261432-01-2

5311217

White to off-white solid powder

1.1±0.1 g/cm3

558.4±50.0 °C at 760 mmHg

174-175°C

291.5±30.1 °C

0.0±1.5 mmHg at 25°C

1.540

5.49

1

7

11

33

805

0

CCOC(=O)C1=C(NC(=C(C1C2=CC=CC=C2/C=C/C(=O)OC(C)(C)C)C(=O)OCC)C)C

GKQPCPXONLDCMU-CCEZHUSRSA-N

InChI=1S/C26H33NO6/c1-8-31-24(29)21-16(3)27-17(4)22(25(30)32-9-2)23(21)19-13-11-10-12-18(19)14-15-20(28)33-26(5,6)7/h10-15,23,27H,8-9H2,1-7H3/b15-14+

diethyl 2,6-dimethyl-4-[2-[(E)-3-[(2-methylpropan-2-yl)oxy]-3-oxoprop-1-enyl]phenyl]-1,4-dihydropyridine-3,5-dicarboxylate

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)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.49 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.

---

Solubility in Formulation 2: ≥ 2.5 mg/mL (5.49 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.

---

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