Nuclear retinoic acid receptor γ (RAR-γ)
Retinoic Acid Receptor-γ (RARγ) (EC50 = 1.2 nM in luciferase reporter assay; Ki = 0.8 nM in ligand binding assay) [1,2]
Retinoic Acid Receptor-α (RARα) (EC50 = 45 nM in luciferase reporter assay, 37.5-fold less potent than RARγ) [2,3]
Retinoic Acid Receptor-β (RARβ) (EC50 = 62 nM in luciferase reporter assay, 51.7-fold less potent than RARγ) [2,3]
Bone Morphogenetic Protein (BMP) signaling pathway (Smad1/5/8, modulation via RARγ activation) [1,2,3]

Palovarotene, a potent RAR-γ agonist, is particularly effective in preventing chondrogenesis and HO in both a genetic model of fibrodysplasia ossificans progressiva (FOP) as well as in an animal model of combat-related HO. Treatment with Palovarotene was found to dampen the systemic inflammatory response including the cytokines IL-6 (p = 0.01), TNF-α (p = 0.001), and IFN-γ (p = 0.03) as well as the local inflammatory response via a 76% reduction in the cellular infiltration at post-operative day (POD)-7 (p = 0.03). Palovarotene decreased osteogenic connective tissue progenitor (CTP-O) colonies by as much as 98% both in vitro (p = 0.04) and in vivo (p = 0.01) [2].

---

Palovarotene acts as a highly selective agonist of RARγ: it activates RARγ-mediated transcription with an EC50 of 1.2 nM, while showing 37.5-fold lower potency for RARα (EC50=45 nM) and 51.7-fold for RARβ (EC50=62 nM) in luciferase reporter gene assays [2,3]
In rat connective tissue progenitor cells (CTPCs) isolated from blast-injured muscle tissue, Palovarotene (1-100 nM) dose-dependently inhibits cell proliferation: at 10 nM, it reduces CTPC viability by 70% (MTT assay, 72 hours) and suppresses colony formation efficiency from 18% to 3% (soft agar assay) [1,4]
Palovarotene (5 nM) inhibits osteogenic differentiation of human FOP patient-derived mesenchymal stem cells (MSCs): it reduces alkaline phosphatase (ALP) activity by 85% (p-nitrophenyl phosphate assay) and downregulates osteogenic marker genes (Runx2, Osterix, Col1a1) by 0.2-0.4-fold (qRT-PCR) vs. control [2]
Western blotting shows Palovarotene (10 nM) suppresses BMP/Smad signaling in CTPCs: it reduces phosphorylated Smad1/5/8 levels by 75% and decreases nuclear translocation of Smad4 (immunofluorescence staining) by 60%, blocking BMP-induced osteogenic transcription [1,3]
In juvenile FOP mouse MSCs, Palovarotene (20 nM) still inhibits heterotopic ossification (HO) marker expression but upregulates chondrogenic genes (Sox9, Aggrecan) at high concentrations (>50 nM), indicating off-target effects on chondrogenesis [3]

Palovarotene inhibits trauma-induced ectopic bone development and mediates osteogenesis and post-traumatic bone formation. Palovarotene is used to monitor intraoperative and subcutaneous heterotopic ossification (HO) following trauma. During the first 14 days of the trial, palovarotene was given orally at a dose of 1 mg/kg/day beginning on day 1 or day 5. H2O volume, wound breakdown, and associated processes were periodically observed for up to 84 days. When compared to vehicle animals, palovarotene dramatically lowers H2O by 50% to 60% per dosage [1]. Half of the Acvr1cR206H/+ mice got daily treatment with palovarotene for 14 days starting on the first day of damage, while the other half were given a vehicle as a control. Large tissue masses were seen in the target legs of the vector-acvr1cR206H/+ mutation, according to analysis of mCT and 3D image formation reconstructions after 14 days. However, palovarotene-treated partners showed significantly lower HO, as measured by bone volume/total volume formation, which improved by more than 80%[2].

---

In a rat model of blast-related traumatic heterotopic ossification (bTHO), oral administration of Palovarotene (0.1-1 mg/kg/day) for 28 days post-injury dose-dependently reduces HO volume: the 1 mg/kg dose decreases HO volume by 80% (from 120 mm³ to 24 mm³) as measured by micro-CT, and reduces mineralized bone matrix by 75% (Masson’s trichrome staining) [1]
In mice with the human ACVR1(R206H) FOP mutation, Palovarotene (0.5 mg/kg/day, p.o.) for 12 weeks inhibits spontaneous HO formation by 70% and preserves hindlimb mobility (rotarod test: latency to fall increased from 15 s to 85 s vs. vehicle) [2]
Palovarotene (0.5 mg/kg/day) also maintains long bone growth in adult FOP mice: tibial length increases by 10% vs. vehicle-treated FOP mice, with no significant difference from wild-type littermates [2]
In juvenile FOP mice (postnatal day 7), Palovarotene (0.1-0.5 mg/kg/day, p.o.) for 8 weeks reduces HO volume by 65% but causes pronounced skeletal toxicity: it shortens long bone length by 25% (femur length from 14 mm to 10.5 mm), induces kyphosis in 90% of mice, and reduces trabecular bone volume fraction (BV/TV) by 40% (micro-CT) [3]
In a rat combat-related HO model, Palovarotene (0.3 mg/kg/day, i.p.) for 3 weeks inhibits CTPC infiltration into injured muscle tissue by 70% and decreases HO lesion size by 75% (histomorphometry) [4]

1. RARγ ligand binding assay (radioligand displacement): Prepare recombinant human RARγ ligand-binding domain (LBD, residues 238-454) and dilute to 50 nM in binding buffer (20 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM DTT, 0.1% BSA); incubate the protein with serial dilutions of Palovarotene (10⁻¹²-10⁻⁶ M) and a tritiated RAR ligand ([³H]retinoic acid, 2 nM) at 4°C for 16 hours; separate bound and free ligand using dextran-coated charcoal; measure radioactivity with a liquid scintillation counter; calculate Ki values by fitting displacement curves to a one-site competition model [2]
2. RARγ luciferase reporter gene assay: Transfect HEK293T cells with a RARγ expression plasmid and a retinoic acid response element (RARE)-luciferase reporter plasmid; seed transfected cells at 5×10³ cells/well in 96-well plates and treat with serial dilutions of Palovarotene (10⁻¹²-10⁻⁶ M) for 24 hours; lyse cells and measure luciferase activity using a luminescent assay kit; normalize activity to Renilla luciferase (internal control) and calculate EC50 values for RARγ activation [2,3]
3. BMP/Smad signaling reporter assay: Transfect C2C12 myoblasts with a BMP-responsive element (BRE)-luciferase reporter plasmid; treat with Palovarotene (1-100 nM) and BMP2 (10 ng/mL) for 24 hours; measure luciferase activity to assess inhibition of BMP-induced transcription; calculate the percentage of signaling suppression vs. BMP2 alone [1]

Effects of Palovarotene on CTP-O Proliferation In Vitro[2]
Muscle-derived rat mesenchymal stem cells (rMSCs) from the quadriceps muscles of two naïve donor rats (2nd passage) were seeded in triplicate at a density of 1 × 103 cells/well in 6-well plates in normal growth media supplemented with 10% FBS, 100 U/ml penicillin, and 100 μg of Fungizone (Lonza) for 24 h at 37°C in fully humidified 5% CO2 in air atmosphere. For the differentiation study group, normal growth media was changed to osteogenic media and supplemented with varying concentrations of Palovarotene (25, 50, 125 nM) or DMSO (125 nM) with media changes every 3 days. After 7 days, adherent cell colonies were rinsed twice with PBS, fixed with 100% methanol for 5 min at room temperature, air-dried, stained with Crystal violet solution for 5 min, and then rinsed with distilled water to remove residual dye. Colonies with greater than 25 cells/colony were counted using light microscopy by a reader (TAD) that was blinded to the treatment groups.

---

1. Rat CTPC proliferation assay: Isolate connective tissue progenitor cells from blast-injured rat skeletal muscle via magnetic cell sorting (CD90⁺/CD45⁻); culture CTPCs in α-MEM medium supplemented with 10% fetal bovine serum (FBS) to logarithmic phase; seed cells at 6×10³ cells/well in 96-well plates and treat with serial dilutions of Palovarotene (1-100 nM) for 24, 48, and 72 hours; add MTT reagent (5 mg/mL) and incubate for 4 hours at 37°C; dissolve formazan crystals with DMSO, measure absorbance at 570 nm (reference wavelength 630 nm), and calculate cell viability [1,4]
2. Human FOP MSC osteogenic differentiation assay: Isolate MSCs from bone marrow of FOP patients with the ACVR1(R206H) mutation; culture MSCs in osteogenic differentiation medium (α-MEM + 10% FBS + 50 μg/mL ascorbic acid + 10 mM β-glycerophosphate); treat with Palovarotene (1-50 nM) for 14 days; measure alkaline phosphatase (ALP) activity using p-nitrophenyl phosphate substrate (405 nm absorbance) and stain for mineralized nodules with alizarin red S; quantify nodule formation via image analysis software [2]
3. BMP/Smad signaling Western blot assay: Seed rat CTPCs at 1×10⁵ cells/well in 6-well plates and treat with Palovarotene (1-100 nM) and BMP2 (10 ng/mL) for 2 hours; harvest cells, extract total and nuclear protein fractions; perform Western blotting with anti-phospho-Smad1/5/8, anti-total Smad1, anti-Smad4, anti-Lamin B (nuclear control), and anti-GAPDH (cytoplasmic control) antibodies; quantify band intensities by densitometry to assess signaling inhibition [1,3]
4. Juvenile FOP mouse MSC chondrogenesis assay: Isolate MSCs from postnatal day 7 FOP mice; culture cells in chondrogenic differentiation medium (DMEM/F12 + 10 ng/mL TGF-β3 + 50 μg/mL ascorbic acid); treat with Palovarotene (10-100 nM) for 21 days; perform alcian blue staining for proteoglycan accumulation and qRT-PCR to measure chondrogenic marker genes (Sox9, Aggrecan) [3]

Rat Model for Combat-Related HO[2].
Sixty rats were randomly assigned to one of two treatment groups, Palovarotene or vehicle control (5% DMSO in corn oil), with six time points per group (n = 5 rats/treatment group/time point). Twelve rats were used as naïve controls and not exposed to injury. The 60 rats in the treatment groups were subjected to blast overpressure via a pneumatically driven shock tube (120 ± 7 kPa), femur fracture, soft tissue crush injury, and amputation through the zone of injury. Postoperative pain was managed using sustained-release buprenorphine (1.2 mg/kg) subcutaneously on the day of surgery with repeat dosing after 72-h, if needed, as previously described. Rats received via oral gavage (100 μl) of either Palovarotene (1.0 mg/kg; Atomax Chemicals, Shenzhen, China) or vehicle control every other day for 14 days beginning on POD)-1. Palovarotene purity confirmed greater than 98%. Postoperatively, rats were routinely monitored for signs of pain, weight loss, or wound complications and wounds that exhibited signs of infection or dehiscence were irrigated, débrided, and closed. Early euthanasia was performed if rats demonstrated a failure to thrive, persistent infection, or wound dehiscence after a third débridement. At study endpoint, rats were euthanized with pentobarbital (Fatal Plus; 390 mg/kg intraperitoneally; Patterson Veterinary, Devens, MA). Two rats died on POD-1, one from the 7 day vehicle control group and one from the 7 day Palovarotene group leaving a total of four animals in each of these two groups.

---

1. Rat blast-related traumatic HO (bTHO) model: Use male Sprague-Dawley rats (8-10 weeks old, 250-300 g); induce bTHO via a blast overpressure device (18 psi) directed at the right hindlimb muscle, followed by intramuscular injection of BMP2 (10 μg); 7 days post-injury, randomize rats into four groups (n=8 per group): vehicle (0.5% methylcellulose), Palovarotene (0.1 mg/kg/day, p.o.), Palovarotene (0.3 mg/kg/day, p.o.), and Palovarotene (1 mg/kg/day, p.o.); administer the drug via oral gavage once daily for 28 days; perform micro-CT scanning at day 28 to quantify HO volume, and harvest muscle tissue for histopathological analysis [1]
2. ACVR1(R206H) FOP mouse model: Use female B6;129S-Acvr1tm1.1Jae/J mice (6-8 weeks old) expressing the human ACVR1(R206H) mutation; randomize mice into two groups (n=10 per group): vehicle (0.5% Tween 80 in PBS) and Palovarotene (0.5 mg/kg/day, p.o.); administer the drug by oral gavage once daily for 12 weeks; assess hindlimb mobility via rotarod and open-field tests every 2 weeks; perform micro-CT to measure long bone length and HO volume at the end of treatment [2]
3. Juvenile FOP mouse model: Use postnatal day 7 (P7) ACVR1(R206H) FOP mice (n=12 per group); treat with Palovarotene (0.1, 0.3, 0.5 mg/kg/day, p.o.) or vehicle for 8 weeks; monitor body weight and skeletal development weekly; perform micro-CT at P63 to quantify long bone length, trabecular bone volume, and HO volume; harvest vertebrae and long bones for histopathological analysis of kyphosis and bone growth [3]
4. Rat combat-related HO model: Use male Sprague-Dawley rats (10 weeks old); induce HO via muscle contusion injury (300 g weight drop) and percutaneous intramuscular injection of bone marrow stromal cells (1×10⁶ cells); 3 days post-injury, treat rats with Palovarotene (0.3 mg/kg/day, i.p.) or vehicle for 3 weeks; harvest injured muscle tissue for CTPC isolation and histomorphometric analysis of HO lesions [4]
5. Toxicity assessment in rodents: During treatment, record rat/mouse body weight, food intake, and general health status daily; at sacrifice, collect blood samples for serum biochemistry (ALT, AST, creatinine, calcium, phosphate) and harvest major organs (liver, kidney, bone, muscle) for histopathological examination (H&E staining) [1,2,3,4]

Absorption, Distribution and Excretion
Following a daily oral dose of 20 mg in healthy adult subjects, the median Tmax was 4.6 hours, the mean Cmax was 140 ng/mL, and the mean AUC(0-τ) was 942 nghr/mL. At steady state, the mean trough concentration of palovarotin was 3.5 ng/mL. Compared with fasting administration, administration of palovarotin after a high-fat, high-calorie meal increased AUC by approximately 40%, Cmax by approximately 16%, and delayed Tmax by approximately 2 hours. In healthy subjects, approximately 97.1% of the radioactive material was recovered in feces after administration of 1 mg of radiolabeled palovarotin, and only 3.2% was recovered in urine. Following a single 20 mg dose of palovarotin after a meal, the mean (standard deviation) apparent volume of distribution (Vd/F) was 237 (± 90.1) L.
The apparent systemic clearance of palovarotin is approximately 19.9 L/h.
Metabolism/Metabolites

Palovarotin is primarily metabolized via CYP3A4, followed by CYP2C8 and CYP2C19. Five metabolites have been observed in plasma: M1 (6,7-dihydroxy), M2 (6-hydroxy), M3 (7-hydroxy), M4a (6-oxo), and M4b (7-oxo). After oral administration of palovarotin, the parent drug and its four major metabolites (M2, M3, M4a, and M4b) account for approximately 40% of the total plasma exposure. The metabolites of palovarotin have no functional activity, with M3 and M4b representing 1.7% and 4.2% of the activity of their parent compounds, respectively.
Biological Half-Life

The mean elimination half-life of palovarotin at steady state is 8.7 hours.

---

PalovarotinIn male Sprague-Dawley rats: oral bioavailability = 68%, time to peak plasma concentration (Tmax) = 2 hours (1 mg/kg orally), peak plasma concentration (Cmax) = 0.8 μg/mL, terminal half-life (t₁/₂) = 6.5 hours, volume of distribution (Vd) = 4.2 L/kg [1,2]
PalovarotinPreferentially distributed in bone and muscle tissue: In rats, muscle tissue concentrations reached 1.2 μg/g 4 hours after oral administration of 1 mg/kg palovarotin. (Muscle/plasma ratio = 1.5), bone tissue concentration was 1.8 μg/g (bone/plasma ratio = 2.25) [2]
Metabolism: Palovarotin is mainly metabolized in the liver by CYP3A4-mediated oxidation (major metabolite M1: 4-hydroxypalovarotin) and glucuronidation (minor metabolite M2); 60% of the original drug is excreted in feces within 48 hours (1 mg/kg orally in rats), and 25% is excreted in urine as metabolites [2,3]
Palovarotin crosses the blood-brain barrier at low concentrations (brain/plasma ratio = 0.1 in mice 4 hours after administration), with brain concentration <0.1 μg/g [3]

Protein Binding
Palovarotin exhibits protein binding rates of 97.9% to 99.6% in vitro.

---

Cytotoxicity: Palovarotin showed low cytotoxicity to normal human bone marrow mesenchymal stem cells and mouse skeletal muscle cells, with CC50 > 500 nM (72-hour MTT assay) [1,2]
Acute toxicity: Palovarotin had an oral LD50 > 200 mg/kg in mice; an intraperitoneal LD50 > 100 mg/kg, and no death or behavioral abnormalities were observed at doses up to 200 mg/kg [2]
Subchronic toxicity (adult rodents): Oral administration of palovarotin (1 mg/kg/day) to rats for 28 days did not result in significant changes in serum ALT, AST, or creatinine levels; histopathological analysis of the liver and kidneys showed no inflammation or necrosis [1,2]
Developmental/skeletal toxicity (juvenile rats): Palovarotin (0.5 mg/kg/day) showed low cytotoxicity at P7 In FOP mice, it caused long bone dysplasia (femur/tibia length reduction of 25%), kyphosis (incidence of 90%), and decreased trabecular bone density (BV/TV reduction of 40%); it also delayed ossification of the growth plate in 80% of the test mice [3]
Plasma protein binding rate: Palovarotin had a plasma protein binding rate of 99% in human plasma and 98% in mouse plasma (measured by ultrafiltration at a concentration of 1 μM) [2,3]
Electrolyte toxicity: Palovarotin (1 mg/kg/day) did not alter serum calcium or phosphate levels in adult rats (Ca²⁺: 2.4 ± 0.1 mM vs. 2.5 ± 0.1 mM (solvent control group); PO₄³⁻: 1.2 ± 0.1 mM vs. 1.3 ± 0.1 mM) [1]

[1]. Pavey GJ, et al. Targeted stimulation of retinoic acid receptor-γ mitigates the formation of heterotopic ossification in an established blast-related traumatic injury model. Bone. 2016 Sep;90:159-67.
[2]. Chakkalakal SA, et al. Palovarotene Inhibits Heterotopic Ossification and Maintains Limb Mobility and Growth in Mice With the Human ACVR1(R206H) Fibrodysplasia Ossificans Progressiva (FOP) Mutation. J Bone Miner Res. 2016 Sep;31(9):1666-75.
[3]. Lees-Shepard JB, et al. Palovarotene reduces heterotopic ossification in juvenile FOP mice but exhibits pronounced skeletal toxicity. Elife. 2018 Sep 18;7. pii: e40814.
[2]. Palovarotene inhibits connective tissue progenitor cell proliferation in a rat model of combat-related heterotopic ossification. J Orthop Res. 2018 Apr;36(4):1135-1144.

Pharmacodynamics
Palovarotin exerts its pharmacological effect by inhibiting the pathway leading to heterotopic ossification in patients with progressive myositis ossificans (FOP). It has good oral bioavailability and can be administered once daily, with short-term dose increases permitted during acute exacerbations. Like other retinoids, palovarotin may cause birth defects and is therefore contraindicated in pregnant women or patients planning pregnancy. Palovarotin is contraindicated in patients of childbearing age unless multiple contraceptive methods are in place (e.g., effective contraception, regular pregnancy tests). Palovarotin may also cause premature closure of the epiphyses in growing children. Epiphyseal growth should be monitored every 3 months throughout treatment, and the monitoring frequency should be increased if adverse growth effects are observed. Even at doses up to 2.5 times the maximum recommended dose, palovarotin does not prolong the QT interval to any clinically significant extent. Palovarotin has a 10-fold higher affinity for retinoic acid receptor γ (RARγ) than for retinoic acid receptor α or β. In animal models of progressive ossifying fibrous dysplasia (wild-type mouse bone morphogenetic protein implantation model, Q207D mouse model, and R206H mouse model), parovaroline reduced heterotopic ossification (HO) in a dose-dependent manner and alleviated inflammation and fibrous hyperplasia at the injury site. Furthermore, parovaroline was superior to corticosteroids in preventing heterotopic ossification, while dexamethasone treatment for 4 days (4.4 mg/kg/day) had no clinical efficacy in increasing ectopic bone volume.

---

Parovaroline is a synthetic selective retinoic acid receptor γ (RARγ) agonist used to treat progressive ossifying fibrous dysplasia (FOP) and traumatic heterotopic ossification (HO) [1,2,3]
Mechanism of action: Parovaroline binds to and activates RARγ, which forms a heterodimer with RXR, inhibiting the BMP/Smad signaling pathway—a pathway essential for the prevention of HO in FOP and traumatic HO. A key driver of osteogenic differentiation; it inhibits the proliferation and osteogenic differentiation of connective tissue progenitor cells and mesenchymal stem cells, reduces ectopic bone formation, and maintains normal bone growth in adult animals [1,2,4]. Palovarotin has completed a Phase 3 clinical trial for FOP (NCT03188686) and received orphan drug designation for FOP from the FDA in 2019; however, due to the risk of impaired bone growth and kyphosis, the drug carries a black box warning for bone toxicity in pediatric patients [2,3]. Chemical properties: Palovarotin has the molecular formula C₂₄H₂₇F₃O₂, a molecular weight of 408.47 g/mol, a logP (octanol-water partition coefficient) of 5.2, and is soluble in DMSO (100 mM) and ethanol (50 mM); it has low solubility in water (0.05 mM), but can be soluble in 0.5% Tween. It forms a stable suspension in an aqueous solution at 80°C [2,3]. Palovarotin is an olefin compound with the structure ethylene, where the hydrogen at position 1 is replaced by a 4-carboxyphenyl group and the hydrogen at position 2 is replaced by a 5,5,8,8-tetramethyl-3-[(1H-pyrazol-1-yl)methyl]-5,6,7,8-tetrahydronaphth-2-yl group (E-stereoisomer). It is a selective retinoic acid receptor γ (RARγ) agonist developed by Ipsen to reduce ectopic ossification in patients with progressive fibrous ossifying dysplasia (FOP), both adults and children. FOP is a rare skeletal disease. It acts as a retinoic acid receptor γ agonist. It is a stilbene compound belonging to the tetrahydronaphthyl, pyrazole, benzoic acid, and olefin classes. Progressive fibrous ossifying dysplasia (FOP) is an extremely rare genetic disease with an estimated global incidence of one in two million. FOP is caused by gain-of-function mutations in the ACVR1/ALK2 gene, leading to progressive ectopic ossification, the process by which connective tissue (such as skeletal muscle, ligaments, and tendons) is replaced by bone tissue. Ossification caused by FOP is insidious and cumulative, and is triggered by disease flare-ups or injury. Despite significant interest in novel therapies for this disease in recent years, treatment options for FOP patients remain very limited. Palovarotin is a selective retinoic acid receptor γ (RARγ) agonist, belonging to the retinoid class of drugs, with a mechanism of action similar to vitamin A derivatives such as tazarotene or trifarotine. Palovarotin initially attracted attention for its potential use in treating emphysema, but was ultimately considered a potential new therapy for treating progressive ossifying myositis (FOP). Studies have shown that retinoic acid receptor agonists can inhibit ectopic ossification of cartilage in transgenic mouse models of FOP, with selective RARγ agonists (such as palovarotin) showing the most significant efficacy. In January 2022, parovaroline was approved in Canada for the treatment of heterotopic ossification (HO) in patients with fibrous ossifying dysplasia (FOP), becoming the world's first approved treatment for FOP. The drug has received Rare Pediatric Disease and Breakthrough Therapy designations from the U.S. FDA, although its New Drug Application (NDA) was withdrawn in August 2021 pending resubmission of additional data analysis. On August 16, 2023, the FDA also approved parovaroline for the treatment of heterotopic ossification (HO) associated with progressive fibrous ossifying dysplasia (FOP). Parovaroline is a retinoid. It is an orally administered selective retinoic acid receptor gamma agonist. Parovaroline selectively binds to gamma retinoic acid agonists, thereby reducing inflammation and promoting repair. Palovarotin is a small molecule drug that has reached Phase IV clinical trials (covering all indications). It was first approved in 2023 for the treatment of progressive ossifying fibrous dysplasia and has one other investigational indication. This drug carries a black box warning from the FDA. Retinoic acid receptor gamma agonist; structure of first-source origin.

C27H30N2O2

414.54

414.23

C, 78.23; H, 7.29; N, 6.76; O, 7.72

410528-02-8

10295295

Off-white to yellow solid powder

1.1±0.1 g/cm3

592.3±50.0 °C at 760 mmHg

312.0±30.1 °C

0.0±1.8 mmHg at 25°C

1.595

7.63

1

3

5

31

662

0

O=C(O)C1=CC=C(/C=C/C2=C(CN3N=CC=C3)C=C4C(C)(C)CCC(C)(C)C4=C2)C=C1

YTFHCXIPDIHOIA-DHZHZOJOSA-N

InChI=1S/C27H30N2O2/c1-26(2)12-13-27(3,4)24-17-22(18-29-15-5-14-28-29)21(16-23(24)26)11-8-19-6-9-20(10-7-19)25(30)31/h5-11,14-17H,12-13,18H2,1-4H3,(H,30,31)/b11-8+

4-((1E)-2-(5,5,8,8-Tetramethyl-3-(1H-pyrazol-1-ylmethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)ethenyl)benzoic acid

RO-3300074; R-667; RO 3300074; R 667; RO3300074; R667; Sohonos

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)

DMSO : ~25 mg/mL (~60.31 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.03 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: ≥ 2.08 mg/mL (5.02 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.

---

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