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.

At all doses, peak plasma concentrations of TBR-652 were reached within 3–4 hours (Table 2). Helmert comparisons showed that steady-state concentrations (Css) were reached on day 8. The area under the concentration-time curve (AUC0–24) and peak plasma concentration (Cmax) after log-normal (ln) transformation at 24 hours indicated that the increase in these parameters was greater than dose-dependent. [1] Figure 5 shows the mean plasma concentrations of TAK-652 at each dose from 30 minutes to 24 hours after administration. At all doses, the drug was detectable in plasma at both 30 minutes and 24 hours after administration. Table 6 lists the estimated pharmacokinetic parameters after a single oral administration in healthy volunteers. Overall, TAK-652 has good oral absorption and a long plasma half-life. At 24 hours after administration of 25 mg, its plasma concentration was 7.2 ng/ml, equivalent to 9.1 nM [5].

Within the studied dose range, TBR-652 was generally safe and well-tolerated. The most common adverse events (AEs) occurring during treatment in subjects receiving the active pharmaceutical ingredient included: gastrointestinal disorders (n = 19, 43%); systemic disorders (n = 11, 25%); neurological disorders (n = 10, 23%); respiratory, thoracic, and mediastinal disorders (n = 10, 23%); infections and parasitic diseases (n = 7, 16%); and mental disorders (n = 5, 11%). Most adverse events in subjects receiving TBR-652 were mild (n = 24) or moderate (n = 5) in severity. Only one subject receiving TBR-652 experienced a serious adverse event, a shoulder abscess, which was determined by the investigator to be a non-serious adverse event unrelated to the study drug, requiring no treatment, and without sequelae. No life-threatening adverse events occurred, the trial was not terminated due to adverse events, and no deaths occurred. All adverse events were determined to be clearly related to the investigational drug. Table 3 lists the adverse events reported as possibly or very likely related to the investigational drug. Most adverse events resolved without intervention. Only 4 subjects in the TBR-652 group required concurrent administration of other medications due to adverse events assessed as possibly related to the investigational drug. These adverse events were: hydrocodone and acetaminophen for abdominal pain in the 25 mg dose group; acetaminophen for headache in the 50 mg dose group; ondansetron for nausea in the 100 mg dose group; and ibuprofen for headache and nausea in one subject in the 150 mg dose group. All adverse events requiring treatment were considered very likely related to the investigational drug. [4]

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No subjects withdrew from the trial due to adverse events among the 24 treated subjects. A total of 4 subjects reported 6 clinical adverse events. Of the 6 adverse events, two dose-independent symptoms (headache and fatigue) were considered to be possibly related to the investigational drug. The remaining four minor adverse events (headache, nasopharyngitis, decreased sensation, and dizziness) appeared to be unrelated to TAK-652 administration, but this conclusion requires further investigation. No treatment- or dose-related trends were observed in serum chemistry, hematology, and urinalysis data during the study. No dose-related trends were also observed in supine systolic blood pressure, diastolic blood pressure, and pulse rate. No treatment- or dose-related trends were observed in vital signs or electrocardiograms of any subject during the study. Of particular note was the absence of evidence of QTc interval prolongation at any dose of TAK-652. No clinically significant changes were observed in electrocardiogram morphology in any subject receiving any dose. No clinically significant changes were observed at the end of the study. Therefore, single oral administration of TAK-652 (25, 50, and 100 mg solutions) is 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 belongs to the benzoxazole octane class of compounds, with the structure (5Z)-1,2,3,4-tetrahydro-1-benzoxazole octane, substituted at positions 1, 5, and 8 with 2-methylpropyl, N-{4-[(S)-(1-propyl-1H-imidazol-5-yl)methylsulfinyl]phenyl}formamido, and 4-(2-butoxyethoxy)phenyl, respectively. It is a potent chemokine 2 and 5 receptor antagonist and is currently under development for the treatment of liver fibrosis caused by non-alcoholic steatohepatitis (NASH) in adults. It also possesses various pharmacological activities, including chemokine 5 receptor antagonist, anti-HIV drug, chemokine 2 receptor antagonist, antirheumatic drug, and anti-inflammatory drug. It is a diether, imidazole compound, sulfoxide, aromatic ether, secondary amide, and benzoxazole octane compound.
Cenicriviroc has been used in research trials for the treatment of HIV infection/AIDS, AIDS-related dementia syndrome, non-alcoholic steatohepatitis, human immunodeficiency virus (HIV), and HIV-1-related cognitive-motor syndrome.
Cenicriviroc is a highly bioavailable, oral dual inhibitor that inhibits human CC chemokine receptors type 2 (CCR2; CD192) and type 5 (CCR5; CD195), possessing potential immunomodulatory, anti-inflammatory, and antiviral activities. After oral administration, Cenicriviroc specifically binds to and blocks the activation of CCR2 and CCR5. This inhibits the CCR2/CCR5-mediated signal transduction pathway and may suppress inflammatory processes. G protein-coupled chemokine receptors CCR2 and CCR5 are expressed on the surface of monocytes and macrophages, stimulating their migration and infiltration; they play a crucial role in inflammation and autoimmune diseases. Furthermore, denicrivirol inhibits the entry of human immunodeficiency virus (HIV)-1 by interacting with the CCR5 co-receptor.
See also: denicrivirol mesylate (note moved to).

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Drug Indications
Treatment of non-alcoholic steatohepatitis (NASH)

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Denicrivirol is a CCR5 antagonist that prevents human immunodeficiency virus type 1 (HIV-1) from entering cells. The CCR5 binding region of the HIV-1 envelope glycoprotein is an important target for neutralizing antibodies (NAbs); therefore, mutations conferring denicrivirol resistance may affect sensitivity to NAbs. This study investigated the relationship between denicriviroc resistance and NAb resistance by in vitro induction of HIV-1 variants resistant to either denicriviroc or neutralizing antibodies (NAbs). The cenicriviroc-resistant variant KK652-67 (KK strain passaged 67 times in the presence of increasing concentrations of cenicriviroc) is sensitive to NAbs targeting the V3 loop, CD4-inducible region (CD4i), and CD4-binding site (CD4bs), while the wild-type (WT) parent HIV-1 strain KKWT, which produced the cenicriviroc-resistant strain KK652-67, is resistant to these NAbs. The V3 region of KK652-67 is crucial for cenicriviroc resistance and is a key factor in the high sensitivity of the V3, CD4i, and CD4bs epitopes to NAbs. Furthermore, inducing a variant resistant to the V3 neutralizing antibody 0.5γ and the CD4i neutralizing antibody 4E9C from the cenicriviroc-resistant strain KK652-67 resulted in a phenotype reverting to that of the parent strain KKWT, which is sensitive to cenicriviroc. Resistance to 0.5γ and 4E9C was caused by novel amino acid substitutions R315K, G324R, and E381K located near the amino acid substitutions that confer resistance to cenicriviroc in the V3 and C3 regions. Importantly, these amino acid changes in the CCR5 binding region also led to a phenotype reverting to the cenicriviroc sensitive phenotype. These results indicate that there are some key amino acid residues on which resistance to cenicriviroc is incompatible with resistance to neutralizing antibodies. This means that cenicriviroc and neutralizing antibodies may limit the emergence of resistant variants of each other. [2]

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The first small molecule CCR5 antagonist, TAK-779, failed to be developed as an anti-human immunodeficiency virus (anti-HIV-1) drug due to its low oral bioavailability. TAK-652 is a derivative of TAK-779 with high oral bioavailability and potent anti-HIV-1 activity. TAK-652 inhibits the binding of RANTES (molecules that activate and regulate the expression and secretion of normal T cells), macrophage inflammatory protein 1α (MIP-1α), and MIP-1β to cells expressing CCR5 at nanomolar concentrations. TAK-652 also inhibits the binding of monocyte chemokine 1 (MCP-1) to cells expressing CCR2b. However, its inhibitory effect on the binding of ligands to other chemokine receptors is limited. TAK-652 is active against (R5) HIV-1 using CCR5 but completely ineffective against (X4) HIV-1 using CXCR4. The compound is active against R5 HIV-1 clinical isolates containing resistance mutations to reverse transcriptase and protease inhibitors, with mean half-maximal effective concentrations (EC50) and EC90s of 0.061 nM and 0.25 nM, respectively. Furthermore, recombinant R5 viruses carrying different subtypes (A to G) of envelope proteins showed similar sensitivity to TAK-652. A single oral dose of up to 100 mg of TAK-652 is safe and well-tolerated in humans. The compound exhibits favorable pharmacokinetic properties, with plasma concentrations remaining as high as 7.2 ng/ml (9.1 nM) even 24 hours after a 25 mg dose. Therefore, TAK-652 is a promising new HIV-1 entry inhibitor. [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.)