CCR5 ( IC50 = 0.29 nM ); CCR2 ( IC50 = 5.9 nM ); R5 HIV-1 ( IC50 = 0.024-0.08 nM ); R5 HIV-2 ( IC50 = 0.03-0.98 nM )

Cenicriviroc (≥20 mg/kg/day) considerably reduces the recruitment of macrophages and monocytes in vivo. In all three animal models of fibrosis, cenicriviroc exhibits antifibrotic effects at these doses, as evidenced by notable decreases in collagen deposition, collagen type 1 protein, and collagen mRNA expression. Cenicriviroc considerably lowers the non-alcoholic fatty liver disease activity score in the NASH model. The weight of the body, liver, or kidneys is not significantly affected by cenicriviroc treatment[1].

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Cenicriviroc (CVC) significantly reduced monocyte/macrophage recruitment in vivo at doses ≥20 mg/kg/day (p < 0.05). At these doses, CVC showed antifibrotic effects, with significant reductions in collagen deposition (p < 0.05), and collagen type 1 protein and mRNA expression across the three animal models of fibrosis. In the NASH model, CVC significantly reduced the non-alcoholic fatty liver disease activity score (p < 0.05 vs. controls). CVC treatment had no notable effect on body or liver/kidney weight. Conclusions: CVC displayed potent anti-inflammatory and antifibrotic activity in a range of animal fibrosis models, supporting human testing for fibrotic diseases. Further experimental studies are needed to clarify the underlying mechanisms of CVC's antifibrotic effects. A Phase 2b study in adults with NASH and liver fibrosis is fully enrolled (CENTAUR Study 652-2-203; NCT02217475). [1]

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Subjects were randomized in a ratio of 4:1 per dose level to Cenicriviroc/TBR-652 (25, 50, 75, 100, or 150 mg) or placebo, taken once daily for 10 days. Changes from baseline in HIV-1 RNA and CD4 cell counts were measured through day 40 and for monocyte chemotactic protein-1 (MCP-1), high-sensitivity C-reactive protein (hs-CRP), and IL-6 at day 10. Pharmacokinetic data were analyzed using noncompartmental statistics. Laboratory and clinical adverse events (AEs) and electrocardiogram changes were recorded. Results: Maximum median reductions in HIV-1 RNA values for the 25, 50, 75, and 150 mg doses were -0.7, -1.6, -1.8, and -1.7 log10 copies per milliliter, respectively. All changes were significant. Median time to nadir was 10-11 days. Suppression persisted well into the posttreatment period. Mean MCP-1 increased significantly by day 10 in the 50-mg and 150-mg dose groups. Effects on CD4 cell counts, hs-CRP, and IL-6 levels were negligible. TBR-652 was generally safe and well tolerated, with no withdrawals due to AEs. Conclusions: TBR-652 caused significant reductions in HIV-1 RNA at all doses. Significant increases in MCP-1 levels suggested a strong CCR2 blockade. TBR-652 was generally well tolerated with no dose-limiting AEs. Pharmacodynamics indicate that TBR-652 warrants further investigation as an unboosted once-daily oral CCR5 antagonist with potentially important CCR2-mediated anti-inflammatory effects.[4]

Cenicriviroc blocks HIV-1 from entering cells at an effective concentration of 50% EC50 of 0.03, 0.33, 0.45, and 0.98 nM for the four R5 HIV-2 clinical isolates that were tested. Cenicriviroc resistance in the dual-tropic and X4-tropic HIV-2 strains is >1000 nM for EC50 and 33% and 4% for MPI, respectively.

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Chemokine binding assay.[5]
The assay procedure for chemokine binding inhibition by test compounds has been described previously. In brief, CCR5-expressing CHO cells were incubated with various concentrations of Cenicriviroc (CVC)/TAK-652 in binding buffer (Ham's F-12 medium containing 20 mM HEPES and 0.5% bovine serum albumin, pH 7.2) containing either 200 pM 125I-regulated on activation, normal T-cell expressed and secreted (RANTES), 125I-macrophage inflammatory protein 1α (MIP-1α), or 125I-MIP-1β. Binding reactions were performed at room temperature for 40 min. The binding reaction was terminated by washing out the cell-free ligand twice with cold phosphate-buffered saline (PBS). The cell-associated radioactivity was recorded with a scintillation counter. Assays of the inhibitory effect of Cenicriviroc (CVC)/TAK-652 on the binding of 125I-RANTES to CCR1, 125I-monocyte chemotactic protein 1 (MCP-1) to CCR2b, 125I-eotaxin to CCR3, 125I-thymus and activation-regulated chemokine (TARC) to CCR4, and 125I-MIP-3β to CCR7 were carried out in a similar manner.

Cenicriviroc phenotypic activity has been tested using a PBMC phenotypic susceptibility assay against four R5-, one X4- and one dual-tropic HIV-2 clinical primary isolates. All isolates were obtained by co-cultivation of PHA-activated PBMC from distinct HIV-2-infected CCR5-antagonist-naïve patients included in the French HIV-2 cohort and were previously tested for maraviroc susceptibility using the same protocol. HIV-2 tropism was determined by phenotypic assay using Ghost(3) cell lines. Results: Regarding the 4 R5 HIV-2 clinical isolates tested, effective concentration 50% EC50 for cenicriviroc were 0.03, 0.33, 0.45 and 0.98 nM, similar to those observed with maraviroc: 1.13, 0.58, 0.48 and 0.68 nM, respectively. Maximum percentages of inhibition (MPI) of cenicriviroc were 94, 94, 93 and 98%, similar to those observed with maraviroc (93, 90, 82, 100%, respectively). The dual- and X4-tropic HIV-2 strains were resistant to cenicriviroc with EC50 >1000 nM and MPI at 33% and 4%, respectively. Conclusions: In this first study assessing HIV-2 susceptibility to cenicriviroc, we observed an in vitro activity against HIV-2 R5-tropic strains similar to that observed with maraviroc. Thus, cenicriviroc may offer a once-daily treatment opportunity in the limited therapeutic arsenal for HIV-2. Clinical studies are warranted.[3]

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Male C57BL/6 mice receive an intraperitoneal injection of TG, and 48 hours later, peritoneal lavage is used to collect activated macrophages. A Transwell1 Chamber with a 5 μm-pore size polycarbonate filter is used to assay chemotaxis. Briefly put, cells are cultured for two hours with 1 nM CCL2 and/or 1 μM Cenicriviroc (dissolved in 0.5% acetic acid dimethyl sulfoxide and diluted 1:1000 with serum-free Roswell Park Memorial Institute-1640 medium and 0.5% bovine serum albumin). Using a 3-laser BD FACSCanto, cells are extracted from the lower compartment and subjected to flow cytometry analysis in order to count the number of F4/80+CD11b+ macrophages. The software FlowJo is used to analyze the results.

Male C57BL/6 mice (n = 44; 8–10 weeks old) are divided into the following groups and given oral gavage (PO) treatments on Days 1–5: non-disease control, twice-daily (BID) vehicle control, 5 mg/kg/day (Cenicriviroc5) BID, 20 mg/kg/day (Cenicriviroc20) BID, 100 mg/kg/day (Cenicriviroc100) BID, Cenicriviroc20 QD, and positive control (1 mg/kg QD)—a corticosteroid that has been shown to reduce inflammation in a number of animal models. On Day 4, all groups except the non-disease controls receive an IP injection of TG 3.85% (1 mL/animal) two hours post-dose to induce peritonitis.

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Monocyte/macrophage recruitment was assessed in vivo in a mouse model of thioglycollate-induced peritonitis. CCL2-induced chemotaxis was evaluated ex vivo on mouse monocytes. CVC's antifibrotic effects were evaluated in a thioacetamide-induced rat model of liver fibrosis and mouse models of diet-induced non-alcoholic steatohepatitis (NASH) and renal fibrosis. Study assessments included body and liver/kidney weight, liver function test, liver/kidney morphology and collagen deposition, fibrogenic gene and protein expression, and pharmacokinetic analyses. [1]

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Single-dose safety and pharmacokinetics in humans.[5]
A double-blind phase I trial was conducted to evaluate the safety, tolerability, and pharmacokinetics of a single oral administration of TAK-652 in humans. Twenty-four healthy volunteers were enrolled in this study (two for a placebo and six for each dose), and three doses (25, 50, and 100 mg) of TAK-652 were administered orally as a solution to individuals in a fasted state. The TAK-652 solution was formulated in 0.5% (wt/vol) methylcellulose with 0.1% (wt/vol) Polysorbate 80 and 2 mM hydrochloric acid in distilled water. The placebo solution was 0.5% (wt/vol) methylcellulose with 0.1% (wt/vol) Polysorbate 80 and 2 mM hydrochloric acid in distilled water. Doses were selected based on allometric scaling of preclinical pharmacokinetic data and considerations of preclinical toxicology (no observed adverse effects). Screening was performed in the 3-week period prior to dosing, and poststudy assessments were carried out at 5 to 7 days postdosing. Safety and tolerability were evaluated by physical examinations (screening and poststudy), recording of vital signs (screening, predose, 1, 2, 4, 8, and 24 h postdose, and poststudy), electrocardiograms (ECG; screening, predose, 2, 6, and 24 h postdose, and poststudy), clinical laboratory evaluations (hematology, serum chemistry, and urinalysis; screening, predose, 24 h postdose, and poststudy), and recording of adverse events (predose, 3, 12, and 24 h postdose, and poststudy). Serial blood samples were collected to determine the plasma concentration of TAK-652. Blood samples were collected prior to drug administration (0 h) and then 0.5, 1, 2, 3, 4, 6, 8, 12, and 24 h after administration. The samples were immediately processed, and the plasma concentration of TAK-652 was quantified by liquid chromatography/tandem mass spectrometry. The lower limit of TAK-652 quantification in plasma was 0.05 ng/ml. Pharmacokinetic parameters were estimated by noncompartmental procedures using WinNonlin, version 3.2, Enterprise. The maximum plasma concentration (Cmax) and time to reach Cmax (Tmax) for each subject were calculated from the measured concentrations. The area under the plasma concentration-time curve from time zero to the last quantifiable concentration (AUC0-tz) for each subject was calculated from the measured concentrations by the trapezoidal rule.

Maximum plasma concentrations of TBR-652 were achieved in 3-4 hours at all doses (Table 2). Helmert contrasts showed that steady-state concentrations (Css) were achieved by day 8. Log-normal (ln) transformed 24-hour area-under-the concentration-time curve (AUC0-24) and maximum plasma concentrations (Cmax) suggested a greater than dose-dependent increase in these parameters. [1]

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The mean plasma concentrations of TAK-652 for each dose from 30 min to 24 h postadministration are shown in Fig. 5. For all doses, the drug was at detectable levels in plasma at 30 min and after 24 h. The estimated pharmacokinetic parameters after single oral administration for healthy volunteers are presented in Table 6. Overall, TAK-652 had good oral absorption and a rather long half-life in plasma. Its plasma concentration at 24 h after a 25-mg administration was 7.2 ng/ml, which corresponds to 9.1 nM [5].

TBR-652 was generally safe and well tolerated at the doses studied. The most commonly encountered treatment-emergent AEs for the subjects on active drug were gastrointestinal disorders (n = 19, 43%); general disorders (n = 11, 25%); nervous system disorders (n = 10, 23%); respiratory, thoracic, and mediastinal disorders (n = 10, 23%); infections and infestations (n = 7, 16%); and psychiatric disorders (n = 5, 11%). Most AEs in subjects on TBR-652 were mild (n = 24) or moderate (n = 5) in intensity. There was only 1 severe AE in a TBR-652-treated subject, an abscess on the shoulder, which was judged as nonserious and unrelated to study drug by the investigator, did not require treatment, and resolved without sequelae. There were no life-threatening AEs, no discontinuations because of an AE, and no deaths. No AEs were judged to be definitely related to study drug. AEs reported as possibly or probably related to study drug are listed in Table 3. Most AEs resolved without intervention. Only 4 subjects on TBR-652 required concomitant medication for an AE assessed as possibly related to study drug. These were hydrocodone plus paracetamol for abdominal pain in the 25-mg dose group, paracetamol for headache in the 50-mg dose group, ondansetron for nausea in the 100-mg dose group, and ibuprofen for a subject with headache and nausea in the 150-mg dose group. No AEs requiring treatment were considered probably related to study drug.[4]

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No withdrawal due to adverse events occurred among the 24 treated subjects. A total of six clinical adverse events were reported for four subjects. Among the six events, two dose-independent symptoms (headache and fatigue) were judged to be possibly related to the study drug. The other four mild events (headache, nasopharyngitis, hypoesthesia, and dizziness) did not seem to be attributable to the administration of TAK-652, yet this conclusion should be confirmed by further studies. No treatment- or dose-related trends in serum chemistry, hematology, and urinalysis data were observed during the study. There were no dose-related trends in supine systolic and diastolic blood pressure and pulse rate. No apparent treatment- or dose-related trends in vital signs or ECG were noted for any subjects during the course of this study. In particular, there was no evidence of a prolongation of the QTc interval at any dose of TAK-652. No clinically important findings were observed in ECG morphology for individuals receiving any dose. No clinically significant changes were noted poststudy. Thus, single oral doses (25, 50, and 100 mg in solution) of TAK-652 were safe and well tolerated in healthy male subjects. [5]

[1]. Antifibrotic Effects of the Dual CCR2/CCR5 Antagonist Cenicriviroc in Animal Models of Liver and Kidney Fibrosis. PLoS One. 2016 Jun 27;11(6):e0158156.

[2]. Incompatible Natures of the HIV-1 Envelope in Resistance to the CCR5 Antagonist Cenicriviroc and to Neutralizing Antibodies. Antimicrob Agents Chemother. 2015 Nov 2;60(1):437-5.

[3]. Cenicriviroc, a Novel CCR5 (R5) and CCR2 Antagonist, Shows In Vitro Activity against R5 Tropic HIV-2 Clinical Isolates. PLoS One. 2015 Aug 6;10(8):e0134904.

[4]. Safety, efficacy, and pharmacokinetics of TBR-652, a CCR5/CCR2 antagonist, in HIV-1-infected, treatment-experienced, CCR5 antagonist-naive subjects. J Acquir Immune Defic Syndr. 2011 Jun 1;57(2):118-25.

[5]. TAK-652 inhibits CCR5-mediated human immunodeficiency virus type 1 infection in vitro and has favorable pharmacokinetics in humans. Antimicrob Agents Chemother. 2005 Nov;49(11):4584-91.

Cenicriviroc is a member of the class of benzazocines that is (5Z)-1,2,3,4-tetrahydro-1-benzazocine which is substituted by a 2-methylpropyl, N-{4-[(S)-(1-propyl-1H-imidazol-5-yl)methanesulfinyl]phenyl}carboxamide and 4-(2-butoxyethoxy)phenyl groups at positions 1, 5 and 8, respectively. It is a potent chemokine 2 and 5 receptor antagonist currently in development for the treatment of liver fibrosis in adults with nonalcoholic steatohepatitis (NASH). It has a role as a chemokine receptor 5 antagonist, an anti-HIV agent, a chemokine receptor 2 antagonist, an antirheumatic drug and an anti-inflammatory agent. It is a diether, a member of imidazoles, a sulfoxide, an aromatic ether, a secondary carboxamide and a benzazocine.
Cenicriviroc has been used in trials studying the treatment of HIV-infection/AIDS, AIDS Dementia Complex, Nonalcoholic Steatohepatitis, Human Immunodeficiency Virus, and HIV-1-Associated Cognitive Motor Complex.
Cenicriviroc is an orally bioavailable, dual inhibitor of human C-C chemokine receptor types 2 (CCR2; CD192) and 5 (CCR5; CD195), with potential immunomodulating, anti-inflammatory and antiviral activities. Upon oral administration, cenicriviroc specifically binds to and prevents the activation of both CCR2 and CCR5. This inhibits the activation of CCR2/CCR5-mediated signal transduction pathways and may inhibit inflammatory processes. The G-protein coupled chemokine receptors CCR2 and CCR5 are expressed on the surface of monocytes and macrophages and stimulate their migration and infiltration; they play key roles in inflammation and autoimmune diseases. In addition, cenicriviroc inhibits human immunodeficiency virus (HIV)-1 entry via CCR5 coreceptor interaction.
See also: Cenicriviroc Mesylate (annotation moved to).

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Drug Indication
Treatment of non-alcoholic steatohepatitis (NASH)

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Cenicriviroc is a CCR5 antagonist which prevents human immunodeficiency virus type 1 (HIV-1) from cellular entry. The CCR5-binding regions of the HIV-1 envelope glycoprotein are important targets for neutralizing antibodies (NAbs), and mutations conferring cenicriviroc resistance may therefore affect sensitivity to NAbs. Here, we used the in vitro induction of HIV-1 variants resistant to cenicriviroc or NAbs to examine the relationship between resistance to cenicriviroc and resistance to NAbs. The cenicriviroc-resistant variant KK652-67 (strain KK passaged 67 times in the presence of increasing concentrations of cenicriviroc) was sensitive to neutralization by NAbs against the V3 loop, the CD4-induced (CD4i) region, and the CD4-binding site (CD4bs), whereas the wild-type (WT) parental HIV-1 strain KKWT from which cenicriviroc-resistant strain KK652-67 was obtained was resistant to these NAbs. The V3 region of KK652-67 was important for cenicriviroc resistance and critical to the high sensitivity of the V3, CD4i, and CD4bs epitopes to NAbs. Moreover, induction of variants resistant to anti-V3 NAb 0.5γ and anti-CD4i NAb 4E9C from cenicriviroc-resistant strain KK652-67 resulted in reversion to the cenicriviroc-sensitive phenotype comparable to that of the parental strain, KKWT. Resistance to 0.5γ and 4E9C was caused by the novel substitutions R315K, G324R, and E381K in the V3 and C3 regions near the substitutions conferring cenicriviroc resistance. Importantly, these amino acid changes in the CCR5-binding region were also responsible for reversion to the cenicriviroc-sensitive phenotype. These results suggest the presence of key amino acid residues where resistance to cenicriviroc is incompatible with resistance to NAbs. This implies that cenicriviroc and neutralizing antibodies may restrict the emergence of variants resistant to each other.[2]

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The first small-molecule CCR5 antagonist, TAK-779, could not be developed as an anti-human immunodeficiency virus type (anti-HIV-1) agent because of its poor oral bioavailability. TAK-652 is an orally bioavailable TAK-779 derivative with potent anti-HIV-1 activity. TAK-652 inhibited the binding of RANTES (regulated on activation, normal T-cell expressed and secreted), macrophage inflammatory protein 1alpha (MIP-1alpha), and MIP-1beta to CCR5-expressing cells at nanomolar concentrations. TAK-652 could also suppress the binding of monocyte chemotactic protein 1 (MCP-1) to CCR2b-expressing cells. However, its inhibitory effect on ligand binding to other chemokine receptors was limited. TAK-652 was active against CCR5-using (R5) HIV-1 but totally inactive against CXCR4-using (X4) HIV-1. The compound was active against R5 HIV-1 clinical isolates containing reverse transcriptase and protease inhibitor-resistant mutations, with a mean 50% effective concentration (EC50) and EC90 of 0.061 and 0.25 nM, respectively. In addition, recombinant R5 viruses carrying different subtype (A to G) envelope proteins were equally susceptible to TAK-652. A single oral administration of TAK-652 up to 100 mg was safe and well tolerated in humans. The compound displayed favorable pharmacokinetics, and its plasma concentration was 7.2 ng/ml (9.1 nM) even 24 h after the administration of 25 mg. Thus, TAK-652 is a promising candidate as a novel entry inhibitor of HIV-1.[5]

C41H52N4O4S

696.94

696.37

C, 70.66; H, 7.52; N, 8.04; O, 9.18; S, 4.60

497223-25-3

Cenicriviroc Mesylate; 497223-28-6; 497223-22-0 (Cenicriviroc Sulfone)

11285792

Light yellow to yellow solid powder

1.2±0.1 g/cm3

913.5±65.0 °C at 760 mmHg

506.3±34.3 °C

0.0±0.3 mmHg at 25°C

1.603

10.22

1

7

17

50

1060

1

[S@](C([H])([H])C1=C([H])N=C([H])N1C([H])([H])C([H])([H])C([H])([H])[H])(C1C([H])=C([H])C(=C([H])C=1[H])N([H])C(C1=C([H])C2C([H])=C(C3C([H])=C([H])C(=C([H])C=3[H])OC([H])([H])C([H])([H])OC([H])([H])C([H])([H])C([H])([H])C([H])([H])[H])C([H])=C([H])C=2N(C([H])([H])C([H])([H])C1([H])[H])C([H])([H])C([H])(C([H])([H])[H])C([H])([H])[H])=O)=O |c:36|

PNDKCRDVVKJPKG-WHERJAGFSA-N

InChI=1S/C41H52N4O4S/c1-5-7-22-48-23-24-49-38-15-10-32(11-16-38)33-12-19-40-35(25-33)26-34(9-8-21-44(40)28-31(3)4)41(46)43-36-13-17-39(18-14-36)50(47)29-37-27-42-30-45(37)20-6-2/h10-19,25-27,30-31H,5-9,20-24,28-29H2,1-4H3,(H,43,46)/b34-26+/t50-/m0/s1

(5E)-8-[4-(2-butoxyethoxy)phenyl]-1-(2-methylpropyl)-N-[4-[(S)-(3-propylimidazol-4-yl)methylsulfinyl]phenyl]-3,4-dihydro-2H-1-benzazocine-5-carboxamide

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: ≥ 2.08 mg/mL (2.98 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.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (2.98 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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