 Chemical Structure.png)
Sufugolix (formerly known as TAK-013) is a novel, highly potent, non-peptide, selective and orally bioavailable luteinizing hormone-releasing hormone (LHRH) receptor antagonist with anticancer activity. It inhibits LHRH with an IC50 of 0.1 nM. Sufugolix was under development for the treatment of endometriosis and uterine leiomyoma and reached phase II clinical trials for both of these indications, but was discontinued.
Physicochemical Properties
| Molecular Formula | C36H31N5O4F2S |
| Molecular Weight | 667.72424 |
| Exact Mass | 667.206 |
| CAS # | 308831-61-0 |
| PubChem CID | 3038517 |
| Appearance | Light yellow to yellow solid powder |
| Density | 1.383g/cm3 |
| Index of Refraction | 1.667 |
| LogP | 6.986 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 10 |
| Heavy Atom Count | 48 |
| Complexity | 1090 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | UCQSBGOFELXYIN-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C36H31F2N5O4S/c1-41(20-23-10-5-3-6-11-23)21-28-31-33(44)43(26-12-7-4-8-13-26)36(46)42(22-27-29(37)14-9-15-30(27)38)34(31)48-32(28)24-16-18-25(19-17-24)39-35(45)40-47-2/h3-19H,20-22H2,1-2H3,(H2,39,40,45) |
| Chemical Name | 1-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difluorobenzyl)-2,4-dioxo-3-phenyl-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-3-methoxyurea |
| Synonyms | TAK013; TAK-013; TAK 013. |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
IC50: 0.1 nM (human LHRH), 0.6 nM (monkey LHRH)[1] Human luteinizing hormone-releasing hormone (LHRH) receptor (IC50 = 0.1 nM in binding assay; IC50 = 0.06 nM in functional antagonism assay) Monkey LHRH receptor (IC50 = 0.6 nM in binding assay; IC50 = 10 nM in functional antagonism assay) Rat LHRH receptor (IC50 > 1000 nM in binding assay) [1] |
| ln Vitro |
Sufugolix shows selectivity for the human receptor over the monkey and rat receptors that is more than three and two thousand times, respectively. On CHO cells expressing the human (IC50=0.1 nM) and monkey (IC50=0.6 nM) receptors, sufugolix effectively inhibits LHRH function. Using high-temperature molecular dynamics computation for the conformational analysis of sufugolix, it is found that the trans conformer of the methoxyurea is less populated than the cis conformer [1]. Sufugolix (TAK-013) exhibited high binding affinity for the cloned human LHRH receptor expressed in Chinese hamster ovary (CHO) cells, with an IC50 value of 0.1 nM in a competitive binding assay using [¹²⁵I]leuprorelin as the radioligand. [1] In a functional assay measuring the inhibition of LHRH-stimulated arachidonic acid (AA) release from CHO cells expressing the human LHRH receptor, Sufugolix (TAK-013) demonstrated potent antagonistic activity with an IC50 value of 0.06 nM. [1] The compound showed species specificity, with significantly lower affinity for monkey (IC50 = 0.6 nM in binding, 10 nM in functional assay) and rat (IC50 > 1000 nM in binding) LHRH receptors compared to the human receptor. [1] |
| ln Vivo |
In castrated male cynomolgus monkeys, oral administration of sufugolix at a dose of 30 mg/kg results in an almost complete suppression of the plasma LH levels with a long enough half-life (more than 24 hours). Sufugolix has maximal plasma concentrations of 0.34 μM at 6 hours after treatment and 0.18 μM at 4 hours after administration at dosages of 30 and 10 mg/kg, respectively[1]. Oral administration of Sufugolix (TAK-013) at a dose of 30 mg/kg to castrated male cynomolgus monkeys caused almost complete suppression of plasma luteinizing hormone (LH) levels (to 11% of pretreatment levels at 24 hours). The suppressive effect lasted for more than 24 hours. [1] At a lower oral dose of 10 mg/kg, Sufugolix (TAK-013) also effectively suppressed plasma LH levels (to approximately 20% of pretreatment levels from 8 to 24 hours post-dose). [1] The in vivo antagonistic effect of Sufugolix (TAK-013) at 30 mg/kg (po) was more potent and had a longer duration of action than the predecessor compound T-98475 at 60 mg/kg (po). [1] |
| Enzyme Assay |
A receptor binding assay was performed to determine the affinity of compounds for the LHRH receptor. Membrane fractions prepared from CHO cells stably expressing the recombinant human LHRH receptor were incubated with a fixed concentration of the radioligand [¹²⁵I]leuprorelin (0.12-0.15 nM) and various concentrations of the test compound at 25°C for 60 minutes in an assay buffer. The reaction was terminated by adding ice-cold buffer, and bound ligand was separated by rapid filtration through a polyethylenimine-coated glass fiber filter. Radioactivity on the filter was measured. Specific binding was calculated by subtracting nonspecific binding (determined in the presence of 1 μM unlabeled leuprorelin) from total binding. IC50 values were derived by fitting the data to a pseudo-Hill equation. [1] Binding assays for monkey and rat LHRH receptors followed similar protocols. For monkey receptors, membranes from CHO cells expressing the cloned cynomolgus monkey LHRH receptor were used. For rat receptors, membranes from the anterior pituitary of male Wistar rats were used, and incubation was performed at 4°C for 90 minutes. [1] |
| Cell Assay |
An in vitro functional antagonism assay was conducted using CHO cells expressing either human or monkey LHRH receptors. Cells were seeded into 24-well plates and cultured. They were then labeled by incubation with [³H]arachidonic acid for one day. After washing, the cells were preincubated with the test compounds at 37°C for 60 minutes. LHRH (1 nM final concentration) was added to stimulate arachidonic acid release, and incubation continued for 40 minutes at 37°C. The radioactivity released into the medium was quantified using a liquid scintillation counter to determine the inhibitory effect of the compounds on LHRH-stimulated response. IC50 values were calculated. [1] |
| Animal Protocol |
Monkeys: Sufugolix (10 or 30 mg/kg, 3 mL/kg, n=3 for each group) is suspended in 0.5% methylcellulose containing 1.2% citric acid, or 0.5% methylcellulose containing 1.2% citric acid alone (3 mL/kg, n=3), are administered orally. Blood samples (heparin-plasma) are collected from a femoral vein 24 h before administration and 0, 2, 4, 8, 24, and 48 h after administration. LH concentrations in the plasma are measured by bioassays using mouse testicular cells[1]. For oral absorption studies, female cynomolgus monkeys (4-8 years old) were used. Compounds (Sufugolix (TAK-013) hydrochloride, 9a, 9g) were suspended in 0.5% methylcellulose and orally administered at a dose of 10 mg/kg. Blood samples (heparinized plasma) were collected from a forearm vein at 1, 3, and 6 hours after administration. Plasma concentrations of the compounds were determined by reverse-phase HPLC. [1] For in vivo efficacy studies evaluating LH suppression, male cynomolgus monkeys (4-9 years old) that had been castrated more than 6 months prior were used. Sufugolix (TAK-013) (free base) was suspended in 0.5% methylcellulose containing 1.2% citric acid and administered orally at doses of 10 or 30 mg/kg (3 mL/kg). A vehicle control (0.5% methylcellulose with 1.2% citric acid) was also administered. Blood samples (heparinized plasma) were collected from a femoral vein 24 hours before and at 0, 2, 4, 8, 24, and 48 hours after administration. Plasma LH concentrations were measured using a bioassay with mouse testicular cells. [1] |
| ADME/Pharmacokinetics |
In cynomolgus monkeys following a single oral dose of 10 mg/kg, Sufugolix (TAK-013) achieved a maximum plasma concentration (Cmax) of 0.2 μM. The time to reach Cmax (Tmax) was 6 hours. The area under the plasma concentration-time curve from 0 to 6 hours (AUC0-6) was 0.85 μM·h. [1] At oral doses of 10 and 30 mg/kg in monkeys, the observed Cmax values were 0.18 μM and 0.34 μM, reached at 4 and 6 hours post-dose, respectively. [1] The oral absorption of Sufugolix (TAK-013) (methoxyurea) in monkeys was superior to its ethylurea analog (9g) at the same 10 mg/kg dose, as indicated by higher Cmax and AUC values. [1] |
| References |
[1]. Discovery of a thieno[2,3-d]pyrimidine-2,4-dione bearing a p-methoxyureidophenyl moiety at the 6-position: a highly potent and orally bioavailable non-peptide antagonist for the human luteinizing hormone-releasing hormone receptor. J Med Chem. 2003 Jan 2;46(1):113-24. |
| Additional Infomation |
Drug Indication Investigated for use/treatment in endometriosis and uterine fibroids. Mechanism of Action Endometriosis is a painful, fertility threatening condition that is a result of tissue similar to that found in the womb lining growing elsewhere in the pelvis. The condition is sex hormone dependent, with gonadotropin releasing hormone (GnRH), a hypothalamic decapeptide amide that plays an important part in the regulation of reproductive processes, being implicated. TAK-013 behaves as an insurmountable antagonist at the human receptor (hGnRHR). It suppresses gonadotropin release. Sufugolix (TAK-013) is a highly potent, orally bioavailable, non-peptide antagonist of the human LHRH receptor. It belongs to a chemical class based on a thieno[2,3-d]pyrimidine-2,4-dione core structure, developed as a superior surrogate for the earlier thienopyridin-4-one core (e.g., T-98475). [1] A key structural feature is the unique p-(3-methoxyureido)phenyl group at the 6-position of the core. Molecular modeling suggests that the methoxyurea moiety forms an intramolecular hydrogen bond between the aniline NH and the methoxy oxygen atom. This bond is proposed to shield hydrogen-bonding groups from solvent, reduce desolvation energy cost, increase apparent lipophilicity, and thereby enhance membrane permeability and oral absorption. [1] Due to its favorable in vitro potency, species selectivity for the human receptor, and improved oral efficacy profile in monkeys, Sufugolix (TAK-013) was selected as a candidate for clinical trials for the potential treatment of sex-hormone-dependent diseases. [1] |
Solubility Data
| Solubility (In Vitro) |
DMSO : ~20 mg/mL (~29.95 mM) H2O : < 0.1 mg/mL |
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.4976 mL | 7.4882 mL | 14.9763 mL | |
| 5 mM | 0.2995 mL | 1.4976 mL | 2.9953 mL | |
| 10 mM | 0.1498 mL | 0.7488 mL | 1.4976 mL |