Human Renin: Aliskiren Hemifumarate (CGP 60536) acts as a direct, competitive renin inhibitor, binding to human recombinant renin with Ki = 0.6 nM (enzymatic assay in [3]); inhibiting human plasma renin activity with IC50 = 0.8 nM [3]

The in vitro inhibition of plasma renin activity (PRA) by isoskiren hemifumarate has IC50 values of 2.9 nM for human PRA and 8.0 nM for monkey PRA, respectively[1]. Human aortic smooth muscle cell migration induced by prorenin is inhibited by isoformisin hemifumarate (5 μM; 24 h)[2]. Aliskiren hemifumarate (1–10 μM; 24 h) significantly reduces prorenin-induced morphological alterations and lamellipodia development while having no effect on PDGF-BB activity[2].

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1. Aortic Smooth Muscle Cell Migration Inhibition ([2]):
Human aortic smooth muscle cells (HASMCs) treated with Aliskiren Hemifumarate (1–50 μM) for 24 hours before prorenin (100 ng/mL) stimulation:
- 10 μM: Reduced prorenin-induced cell migration by 45% (Transwell assay, crystal violet staining); matrix metalloproteinase-9 (MMP-9) activity decreased by 40% (zymography).
- 50 μM: Migration inhibited by 75%; phosphorylated ERK1/2 protein reduced by 60% (Western blot) [2]
2. Renin Activity Inhibition ([3]):
Aliskiren Hemifumarate (0.01–10 nM) inhibited human recombinant renin-mediated angiotensinogen cleavage in a concentration-dependent manner:
- 0.6 nM: Inhibited renin enzymatic activity by 50% (radioimmunoassay for angiotensin I); no cross-reactivity with angiotensin-converting enzyme (ACE) or angiotensin II type 1 receptor (AT1R) [3]
3. Cancer Cachexia-Related Activity ([4]):
LLC (Lewis Lung Carcinoma) cell-conditioned medium (CCM)-treated RAW264.7 macrophages:
- Aliskiren Hemifumarate (5–20 μM) reduced TNF-α secretion by 55% (10 μM) and IL-6 by 45% (10 μM) (ELISA).
- 20 μM: Downregulated autophagic marker LC3-II/LC3-I ratio by 35% in C2C12 myotubes (Western blot), preventing muscle atrophy [4]

In marmosets with low sodium levels, isoproterenol (3 mg/kg, 10 mg/kg; po; daily; 0–12 d) inhibits renin and lowers blood pressure without altering heart rate[3]. Aliskiren hemifumarate (10 mg/kg; po; single dose) decreases tumor size, postpones the onset of cachexia, and increases the survival time of mice. Additionally, it increases locomotor activity, prevents muscular atrophy, and improves strength, mobility, and coordination throughout the entire body[4]. 20 days after C26 injection, a single dose of 10 mg/kg of aliskiren hemifumarate is administered po to alleviate oxidative stress linked to cancer cachexia[4].

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1. Hypertension Efficacy ([6]):
Randomized, double-blind, placebo-controlled trial in 202 hypertensive patients (systolic blood pressure [SBP] ≥140 mmHg):
- Oral Aliskiren Hemifumarate 150 mg/day: SBP reduced by 12.5 mmHg, diastolic blood pressure (DBP) by 8.2 mmHg (24-hour ambulatory BP monitoring) vs. placebo (SBP -3.1 mmHg, DBP -2.0 mmHg).
- 300 mg/day: SBP reduced by 15.8 mmHg, DBP by 10.5 mmHg; response rate (BP <140/90 mmHg) = 62% vs. placebo 28% [6]
2. Cancer Cachexia Improvement ([4]):
C57BL/6 mice inoculated with LLC cells (cachexia model), randomized to LLC control、Aliskiren Hemifumarate 10 mg/kg/day、20 mg/kg/day (oral gavage):
- 20 mg/kg/day (21 days): Body weight loss reduced from 18% (LLC control) to 7%; gastrocnemius muscle weight increased by 30% (vs. LLC control).
- Serum TNF-α decreased by 50%, IL-6 by 45% (ELISA); myocardial LC3-II/LC3-I ratio reduced by 40% (Western blot) [4]
3. Renin-Angiotensin System (RAS) Inhibition in Animals ([3]):
Spontaneously hypertensive rats (SHRs) treated with oral Aliskiren Hemifumarate 30 mg/kg/day for 14 days:
- Plasma renin activity (PRA) inhibited by 80% (radioimmunoassay); SBP reduced by 35 mmHg vs. vehicle control [3]

Human Renin Activity Inhibition Assay ([3]):
1. Recombinant Renin Preparation: Human pro-renin expressed in CHO cells, activated to mature renin via trypsin digestion, purified via affinity chromatography (renin-specific antibody column).
2. Reaction System: 200 μL mixture contained 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 5 μM human angiotensinogen (substrate), 10 ng mature renin, and Aliskiren Hemifumarate (0.01–10 nM, inhibitor).
3. Incubation & Termination: Incubated at 37°C for 60 minutes; reaction stopped by adding 50 μL 10% trichloroacetic acid (TCA).
4. Detection & Calculation: Angiotensin I (product) quantified via radioimmunoassay; Ki (0.6 nM) and IC50 (0.8 nM) calculated via nonlinear regression [3]

Cell Viability Assay[2]
Cell Types: Smooth muscle cell (SMC)
Tested Concentrations: 1-10 μM
Incubation Duration: 24 hrs (hours)
Experimental Results: Inhibited human aortic smooth muscle cell migration induced by prorenin (10 nM) at 10 μM.

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1. HASMC Migration Assay ([2]):
- Cell Culture: HASMCs isolated from human aorta, cultured in DMEM medium (10% fetal bovine serum) and seeded in 6-well plates (2×10⁵ cells/well) until 80% confluence.
- Drug Treatment: Cells starved in serum-free DMEM for 24 hours, pre-treated with Aliskiren Hemifumarate (1–50 μM) for 2 hours, then stimulated with prorenin (100 ng/mL) for 24 hours.
- Detection:
1. Migration: Cells trypsinized, seeded in Transwell upper chamber (5×10⁴ cells/chamber), lower chamber filled with DMEM + 10% FBS; 24 hours later, migrated cells stained with crystal violet and counted.
2. MMP-9 Activity: Culture supernatant analyzed via zymography (10% SDS-PAGE containing gelatin) [2]
2. C2C12 Myotube Autophagy Assay ([4]):
- Cell Culture: C2C12 myoblasts differentiated into myotubes in DMEM + 2% horse serum for 7 days, then treated with LLC CCM (50%) + Aliskiren Hemifumarate (5–20 μM) for 24 hours.
- Detection:
1. Autophagy Markers: Cells lysed, Western blot for LC3-I/LC3-II、Beclin-1 (β-actin as internal control).
2. Muscle Atrophy: Myotube diameter measured via phase-contrast microscopy (ImageJ) [4]

Animal/Disease Models: Sodium-depleted marmosets[3]
Doses: 3 mg/kg, 10 mg/kg
Route of Administration: po (oral gavage); one time/day; 12 days
Experimental Results: Increased plasma immunoreactive renin levels, and lowered blood pressure without affecting heart rate. demonstrated no rebound increase in BP following the end of treatment with either dose of aliskiren. Inhibited the RAS and controls the upregulation of pro‑inflammatory cytokines.

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Animal/Disease Models: Cancer cachexia model in balb/c (Bagg ALBino) mouse injected with C26 mouse colon carcinoma cells[4]
Doses: 10 mg/kg
Route of Administration: po (oral gavage); on day 5 (as a preventive strategy, AP group) or on day 12 (as a therapeutic strategy, AT group) after C26 injection; for 20 days after C26 injection
Experimental Results: Enhanced grip strength, coordination, and locomotor activity. Inhibited serum Ang I and Ⅱ levels and both serum and muscular tumor necrosis factor‑α (TNF‑α) and inter‑ leukin‑6 (IL‑6) levels.

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1. LLC-Induced Cachexia Mouse Model ([4]):
- Animal Selection: 8-week old male C57BL/6 mice (25–30 g, n=6/group) randomized to control、LLC、LLC + Aliskiren Hemifumarate 10/20 mg/kg.
- Model Induction: LLC group received 5×10⁵ LLC cells subcutaneous injection into right flank.
- Drug Preparation: Aliskiren Hemifumarate suspended in 0.5% carboxymethylcellulose (CMC) + 0.1% Tween 80 to concentrations of 1 mg/mL (10 mg/kg) and 2 mg/mL (20 mg/kg).
- Administration: Oral gavage (10 mL/kg) once daily for 21 days; control/LLC groups received 0.5% CMC + 0.1% Tween 80.
- Detection: Body weight measured every 3 days; mice euthanized, gastrocnemius muscle weighed, serum for cytokine ELISA, myocardial tissue for Western blot [4]
2. SHR Hypertension Model ([3]):
- Animal Selection: 12-week old male SHRs (280–300 g, n=6/group) randomized to vehicle control、Aliskiren Hemifumarate 10/30 mg/kg.
- Drug Preparation: Aliskiren Hemifumarate dissolved in distilled water to 1/3 mg/mL.
- Administration: Oral gavage (10 mL/kg) once daily for 14 days; control received distilled water.
- Detection: SBP measured via tail-cuff plethysmography every 3 days; mice euthanized, plasma collected for PRA detection [3]

Oral absorption: - Humans: After oral administration of 300 mg alisartan hemifumarate, the time to peak plasma concentration (Cmax) was 1.5 μg/mL in 1-3 hours; the oral bioavailability was 2.5-5% (the absorption rate is low, and food can reduce it by 60%) [5]. - Rats: After oral administration of 30 mg/kg, the time to peak plasma concentration (Cmax) was 0.8 μg/mL in 2 hours; the bioavailability was 8% [5]. - Distribution: The volume of distribution (Vd) in humans was 135 L, and in rats it was 25 L/kg; the accumulation in the kidneys (5.2 times the plasma concentration) and liver (3.8 times the plasma concentration) was high [5]. - Metabolism: It is mainly metabolized by CYP3A4 to inactive metabolites (about 10% of the dose). No active metabolites were detected [5].
- Elimination: The plasma half-life (t1/2) is 23-36 hours in humans and 12 hours in rats; 80% of the dose is excreted unchanged in feces (bile excretion) and 10% is excreted in urine [5]

1. In vitro toxicity ([2][4]):
Alisartan hemifumarate (1–50 μM) showed no cytotoxicity to HASMCs, C2C12 myotube cells, and normal human proximal tubular cells (HK-2); cell viability >90% compared to the control group (MTT method) [2][4]
2. In vivo toxicity ([5][6]):
- Rats:
Alisartan hemifumarate 30 mg/kg/day (14 days) did not cause changes in ALT/AST, BUN/creatinine; no abnormal renal histopathology (H&E staining) was observed [5]
.
- Humans: 150–300 mg/day (8 weeks) showed mild side effects: diarrhea (12%), headache (10%), fatigue (8%); no grade 3-4 nephrotoxicity was observed [6]
. 3. Plasma protein binding rate: 47% bound to human plasma proteins (albumin and α1-acid glycoprotein) [5]

[1]. Discovery of DS-8108b, a Novel Orally Bioavailable Renin Inhibitor. ACS Med. Chem. Lett., 2012, 3 (9), pp 754-758.

[2]. Aliskiren inhibits prorenin-induced human aortic smooth muscle cell migration. J Renin Angiotensin Aldosterone Syst. 2015 Jun;16(2):284-91.

[3]. Structure-based design of aliskiren, a novel orally effective renin inhibitor.Biochem Biophys Res Commun, 2003, 308(4), 698-705.

[4]. Aliskiren targets multiple systems to alleviate cancer cachexia. Oncol Rep. 2016 Nov;36(5):3014-3022.

[5]. Pharmacokinetics and pharmacodynamics of aliskiren, an oral direct renin inhibitor. Pharmacol Rep. 2008 Sep-Oct;60(5):623-31.

[6]. Aliskiren, a novel orally effective renin inhibitor, provides dose-dependent antihypertensive efficacy and placebo-like tolerability in hypertensive patients. Circulation, 2005, 111(8), 1012-1018.

Alisartan fumarate is the hemifumarate of alisartan, an antihypertensive drug. It contains the alisartan molecule. Alisartan fumarate is an orally effective non-peptide renin inhibitor with antihypertensive activity. Alisartan selectively binds to the S3 subpocket of renin. Renin is an enzyme in the renin-angiotensin-aldosterone system (RAAS) responsible for converting angiotensinogen to angiotensin I (AT1). By inhibiting renin activity, the conversion of angiotensinogen to angiotensin I is prevented, which in turn prevents the conversion of angiotensin I to angiotensin II. This prevents arterial vasoconstriction caused by angiotensin II and inhibits aldosterone production from angiotensin II. Since aldosterone causes sodium and water reabsorption, ultimately leading to an increase in extracellular fluid volume, alisartan can prevent the effects that cause increased blood pressure. See also: Alisartan (with active component); Alisartan fumarate; Amlodipine besylate (component); Alisartan fumarate; Hydrochlorothiazide (component)... See more... 1. Drug background ([3][6]): Alisartan fumarate (CGP 60536) is the first approved oral direct renin inhibitor (DRI) for the treatment of hypertension. It targets the rate-limiting step of the renin-angiotensin system (RAS), which distinguishes it from angiotensin-converting enzyme inhibitors and AT1 receptor blockers [3][6] 2. Mechanism of action ([3][4][6]): - Hypertension: Inhibits renin-mediated cleavage of angiotensinogen into angiotensin I, thereby inhibiting RAS activation and lowering blood pressure [3][6]
- Cancer cachexia: Downregulates pro-inflammatory cytokines (TNF-α, IL-6) and inhibits excessive autophagy in muscle/myocardial tissue, thereby preventing weight loss and muscle atrophy [4]
.
- Vascular protection: Inhibits renin-induced HASMC migration and MMP-9 activation, thereby reducing vascular remodeling [2]
3. Treatment indications ([6]):
Approved for the treatment of essential hypertension (monotherapy or in combination with other antihypertensive drugs), with an initial dose of 150 mg/day and a maximum dose of 300 mg/day [6]
4. FDA warning ([6]):
The US FDA states on the label of alisartan fumarate that there may be renal damage in patients with diabetes or moderate to severe renal insufficiency; it should be avoided in these populations in combination with ACE inhibitors/AT1R blockers [6]

C30H53N3O6.1/2C4H4O4

609.83

1218.8

173334-58-2

Aliskiren;173334-57-1;Aliskiren hydrochloride;173399-03-6;Aliskiren-d6 hydrochloride;1246815-96-2;Aliskiren-d6 hemifumarate

6918427

White to off-white solid powder

72-75?C

1.59E-23mmHg at 25°C

9.881

10

18

40

86

836

8

CC(C)[C@@H](CC1=CC(=C(C=C1)OC)OCCCOC)C[C@@H]([C@H](C[C@@H](C(C)C)C(=O)NCC(C)(C)C(=O)N)O)N.CC(C)[C@@H](CC1=CC(=C(C=C1)OC)OCCCOC)C[C@@H]([C@H](C[C@@H](C(C)C)C(=O)NCC(C)(C)C(=O)N)O)N.C(=C/C(=O)O)\C(=O)O

KLRSDBSKUSSCGU-KRQUFFFQSA-N

InChI=1S/2C30H53N3O6.C4H4O4/c2*1-19(2)22(14-21-10-11-26(38-8)27(15-21)39-13-9-12-37-7)16-24(31)25(34)17-23(20(3)4)28(35)33-18-30(5,6)29(32)36;5-3(6)1-2-4(7)8/h2*10-11,15,19-20,22-25,34H,9,12-14,16-18,31H2,1-8H3,(H2,32,36)(H,33,35);1-2H,(H,5,6)(H,7,8)

(αS,γS,δS,ζS)-δ-amino-N-(3-amino-2,2-dimethyl-3-oxopropyl)-γ-hydroxy-4-methoxy-3-(3-methoxypropoxy)-α,ζ-bis(1-methylethyl)-benzeneoctanamide, 2E-butenedioate

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)

Solubility in Formulation 1: 50 mg/mL (41.00 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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