NK-3 RECEPTOR ANTAGONISTS FOR THERAPEUTIC OR COSMETIC TREATMENT OF EXCESS BODY FAT

Abstract

Disclosed is use of NK3R antagonists for the therapeutic treatment leptin-related disease.

Claims

1. A method of treatment of a leptin-related disease comprising the administration of an NK-3 receptor antagonist to a patient in need thereof, wherein the NK-3 receptor antagonist is of general formula III: ##STR00079## or a pharmaceutically acceptable solvate thereof, wherein: R.sup.1 is H, F or methyl; R.sup.1′ is H; R.sup.2 is H, F, Cl or methoxy; R.sup.2′ is H or F; R.sup.3 is H, F, Cl, methyl, trifluoromethyl, nitrile or R.sup.3 is thiophen-2-yl under the condition that R.sup.5 is not methyl; R.sup.4 is methyl, ethyl, n-propyl, hydroxyethyl, methoxyethyl, trifluoromethyl, difluoromethyl or fluoromethyl; R.sup.5 is methyl, ethyl, methoxymethyl, trifluoromethyl, difluoromethyl, fluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl or 2,2,2-trifluoroethyl; X.sup.1 is N and X.sup.2 is S or 0; or X.sup.1 is S and X.sup.2 is N; represents a single or a double bound depending on X.sup.1 and X.sup.2; and stands for the (R)-enantiomer or for the racemate compound of formula III.

2. The method according to claim 1, wherein the leptin-related disease is selected from the group consisting of metabolic disorders; lipid regulation disorders; Congenital Leptin Deficiency; hypothalamic amenorrhea; Rabson-Mendenhall syndrome; and osteoporosis.

3. The method according to claim 1, wherein the leptin-related disease is selected from the group consisting of diabetes, cardiovascular diseases, and metabolic syndrome.

4. The method according to claim 1, wherein the leptin-related disease is selected from the group consisting of lipodystrophy, dyslipidemia, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, and hyperlipidemia.

5. The method according to claim 1, wherein the leptin-related disease is selected from the group consisting of congenital lipodystrophy, acquired lipodystrophy, and exercise-induced hypothalamic amenorrhea.

6. The method according to claim 1, wherein the NK-3 receptor antagonist is administrated under the form of a pharmaceutical composition comprising the NK-3 receptor antagonist and at least one pharmaceutically acceptable vehicle.

7. The method according to claim 1, wherein the NK-3 receptor antagonist is (R)-(4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone, or a pharmaceutically acceptable solvate thereof.

8. The method according to claim 1, wherein the NK-3 receptor antagonist is (R)-(4-fluorophenyl)(8-methyl-3-(3-methyl-1,2,4-thiadiazol-5-yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0307] FIG. 1 is a histogram showing the % Fat/Total Mass ratio in the pre-dose phase (left column) versus final week (week-13) of the dosing phase (right column) for each group (vehicle-control group versus treated groups with 10 and 50 mg/kg of compound k-5). Data are presented as mean ±SEM; N=4-6/group, stats by 2-way ANOVA, Bonferroni' s MCT.

[0308] FIG. 2 is a graph showing the plasma leptin level along treatment for the vehicle-control group versus treated group with 50 mg/kg of compound k-5 (*: p<0.05, 2-way ANOVA & Sidak's MCT).

[0309] FIG. 3 is a graph showing the percentage of body weight increase along treatment for the vehicle-control group versus treated group with 50 mg/kg of compound k-5.

[0310] FIG. 4 is a diagram showing plasma leptin level in the pre-dose phase versus 4 h after final dose, for each group (placebo group versus treated groups with 20, 60 and 180 mg of compound k-5) (* p<0.05; paired t-test).

[0311] FIG. 5 is a histogram showing plasma leptin level in baseline estrous cycle (left column) versus treatment estrous cycle (right column) for each group (vehicle-control group versus treated group with 10 mg/Kg of compound k-5) (* p<0.05, paired t-test).

EXAMPLES

[0312] The present invention will be better understood with reference to the following examples.

Example 1: Decrease of % Fat/Total Mass in Monkeys

[0313] The cynomolgus monkey was selected as a relevant species because of the similarity of nonhuman primates (NHP) to humans. The route of administration is oral consistent with the intended route in humans.

[0314] Sexually-mature (age range of ˜4 years at pre-dose), purpose-bred cynomolgus monkeys (Macaca fascicularis) of Asian origin were used. Sexual maturity was proven by recording of at least two menstrual bleedings (with 20-to-50 days between menstruations) prior to entry into the study.

[0315] NHPs were maintained on a standard lab diet of twice-daily offerings of commercial pellets for primates supplemented with fresh fruit and bread. Dietary and environmental enrichment were also provided through the study duration. Animals were housed in a climate (19-25° C.) and humidity (40-70%) controlled environment with artificial lighting controlled automatically to give a cycle of 12 hours light and 12 hours dark.

[0316] NHPs were divided into three treatment groups: vehicle control (N=6) and compound k-5 Dose Groups of 10 mg/kg (N=4) and 50 mg/kg (N=6), respectively. All NHPs were dosed once daily (morning) by oral gavage over the 13 week treatment period of the study. The dose vehicle was 0.5% methylcellulose (Methocel® from Colorcon) in water.

[0317] All NHPs were subjected to a body weight measurement and DEXA scan (Dual-energy X-ray absorptiometry, Hologic Dexa QDR® 4000) in the pre-dose phase and in the final week (week-13) of the dosing phase. The measurements were made under ketamine anesthesia with dorbene and antisedan.

[0318] The whole body was scanned by DEXA for measurement of lean mass, bone mineral density, bone mineral content and fat mass.

[0319] All NHPs were subjected to leptin levels measurements. Blood samples were collected at 08 h00 in the morning from overnight-fasted animals on Day-2 prior to dosing (‘pre-dose’), 24 h after the initial dosing, on day 52 of dosing and on day 86 of dosing. Serum samples were derived from these blood samples and stored frozen (−20° C.) until analysis. Leptin concentrations in these serum samples were determined by ELISA assay (Monkey Kit: MyBiosource Cat. N° MBS705354) based upon linear regression analysis against a standard curve.

[0320] Effect on body weight: A mean weight gain of about 12% was observed in vehicle-treated monkeys over the course of the dosing phase of the study (FIG. 3). In comparison, there was no significant change in mean weight for the k-5 treated group over the course of the dosing phase of the study.

[0321] Effects on body composition: There were no significant changes in lean mass, nor bone mineral density not bone mineral content at any measurement, for any group, over the course of the study. DEXA analysis revealed that % fat/total mass increased in the vehicle group over the 13-week study course, as is often observed as NHPs are confined to study. In contrast, no significant change in % fat/total mass was observed in any of the compound k-5 dose groups over the 13-week treatment period, as presented in FIG. 1. A statistically-significant difference was determined between the vehicle-treated and 50 mg/kg-treated groups where N values were 6 subjects/group. In total, these data demonstrate that treatment with compound k-5 prevents adiposity gain in monkeys while not decreasing lean mass.

[0322] Effects on leptin: FIG. 2 illustrates that compound k-5 treatment significantly increases serum leptin levels 24-hours after initial dosing relative to (a) pre-dose levels within the same group and (b) time-matched levels within the vehicle-treated group. Also, the figure illustrates that serum leptin levels were persistently higher in the k-5-treated group relative to the vehicle-treated group over the duration of dosing.

[0323] Study conclusions: Sexually-mature, female monkeys treated with compounds of the invention have decreased weight gain relative to vehicle treated controls. This difference in weight gain is due entirely to a drug-related reduction in % fat mass relative to vehicle controls and not due to changes in bone or muscle density. Moreover, this decrease in % fat mass, relative to vehicle group, correlates with a significant increase in circulating leptin concentrations in response to treatment.

Example 2: Decrease of Total Body Weight in Healthy Women

Multiple Ascending Dose Administration in Healthy Women

[0324] Three Panels A, B, and C, each consisting of 6 healthy women, were randomized to receive different dose level of compound k-5. In those 3 panels, compound k-5 is administered for 21 consecutive days.

[0325] Subjects were residential from the day before first dosing (D-1) until Day 24 (=72 hours after the last dose intake on Day 21). Weekly non-residential visits were organized on Day 28, Day 35 (±1 day) and a follow-up visit on Day 42 (±2 days). Ascending multiple doses of compound k-5 were administered in a once-daily regimen for 21 consecutive days. The suggested dose levels were: 20 mg, 60 mg and 180 mg based on previous studies. Subjects received the trial medication in a q.d. regimen and after a light breakfast.

Main Criteria for Subject Inclusion

[0326] Subjects were recruited on the basis of their medical history and health status. Especially, subjects meeting all of the following criteria were eligible to participate in this study:

[0327] 1) Premenopausal females between 18 and 45 years of age inclusive.

[0328] 2) Healthy with no clinically significant abnormalities as determined by medical history, physical examination, blood chemistry assessments, hematologic assessments, coagulation and urinalysis, measurement of vital signs, and ECG. Isolated out-of-range values judged by the physician to be of no clinical significance were allowed. This determination had to be recorded in the subject's source documents.

[0329] 3) Had a body weight in the range of 50 to 100 kg inclusive.

[0330] 4) Agreed to abstain from alcohol intake 24 hours before administration of study compound, during the period of the study and 24 hours prior to all other clinic visits.

[0331] 5) Agreed not to use prescription medications within 14 days prior to study compound administration and through the duration of the study, unless approved by the Investigator and Sponsor medical monitor.

[0332] 6) Agreed not to use over-the-counter (OTC) medications (including corticosteroids, aspirin, decongestants, antihistamines, and other non-steroidal anti-inflammatory drug [NSAIDs]), and herbal medication (including herbal tea, St. John's Wort), within 14 days prior to study compound administration through the final follow-up visit, unless approved by the investigator and Sponsor medical monitor. Occasional use of paracetamol at recommended doses was allowed.

[0333] 7) Subjects had to have signed an informed consent document indicating that they understood the purpose of and procedures required for the study and were willing to participate in the study.

[0334] 8) Willing/able to adhere to the study visit schedule and other requirements, prohibitions and restrictions specified in this protocol.

Weight Measurement

[0335] The weight was measured at screening, which took place between 28 and 2 days before start of dosing (D-28 to D-2), and at the end-of-study visit (D42, i.e. 21 days after cessation of study compound intake).

Results

[0336] A two-tailed T-test for paired samples was used to explore the difference between the screening and post-study values for weight. Mean values per dose group are tabulated in the following table:

TABLE-US-00002 Weight Weight Pre vs Post Pre vs Post Screening D 42 Difference Difference Matching Dose group (kg) (kg) (kg) (%) p-value Cpd k-5 20 mg 58.2 58.0 −0.13 −0.23% 0.70 Cpd k-5 60 mg 73.3 71.9 −1.38 −1.92% 0.15 Cpd k-5 180 mg 69.0 66.9 −2.10 −3.14% <0.02

[0337] Based on these results it can be concluded that a dose-dependent trend exists towards weight lowering with increasing doses of compound k-5, reaching a significance level of 0.02 in the highest dose group tested in healthy women.

Example 3: Decrease of Body Weight in Healthy Women with Normal Corpulence

[0338] Healthy women with normal corpulence, i.e. having a BMI ranging from 18.5 to 25 kg/m.sup.2, received 60 mg of compound k-5 for 21 consecutive days.

[0339] The weight was measured at screening which took place between 28 and 2 days before start of dosing (D-28 to D-2), as well as on the follow-up visit planned on D42 (21 days after cessation of study compound intake).

[0340] Weight and BMI before and after dosing are presented below:

TABLE-US-00003 Weight Weight Height Screening D 42 Difference Difference BMI BMI Subject (cm) (kg) (kg) (kg) (%) screening D 42 1 173 66.4 64.2 −2.2 −3.43% 22.18 21.45 2 163 63 61 −2 −3.28% 23.71 22.95 3 178 68.3 67.8 −0.5 −0.74% 21.55 21.39

[0341] These results evidence that the use of a NK3R antagonist enables lowering weight in healthy women.

Example 4: Decrease of Body Weight in Overweight Women

[0342] Overweight women, i.e. having a BMI ranging from 25 to 30 kg/m.sup.2, received 180 mg of compound k-5 for 21 consecutive days.

[0343] The weight was measured at screening which took place between 28 and 2 days before start of dosing (D-28 to D-2), as well as on the follow-up visit planned on D42 (21 days after cessation of study compound intake).

[0344] Weight and BMI before and after dosing are presented below:

TABLE-US-00004 Weight Weight Height Screening D 42 Difference Difference BMI BMI Subject (cm) (kg) (kg) (kg) (%) screening D 42 4 156 68 65.2 −2.8 −4.29% 27.94 26.79 5 160 71.2 67 −4.2 −6.27% 27.81 26.17 6 179 87 84 −3 −3.57% 27.15 26.21

[0345] These results evidence that the use of a NK3R antagonist enables lowering weight in overweight women.

Example 5: Effect on Plasma Leptin Level in Women

[0346] This was a randomized, double-blind, placebo-controlled study. All subjects provided written informed consent prior to screening for study eligibility. Healthy female volunteers aged 20-45 years with a body mass index of 19-30 kg/m.sup.2 were included. Participants needed to be in good physical health including the presence of a regular ovulatory menstrual cycle and the discontinuation of all hormonal contraceptive methods at least 3 months prior to screening. Three panels of 8 non-obese female volunteers (median BMI of each panel: 22.3, 23.4, 25.2 kg/m.sup.2) each were administered capsules of compound k-5 or matching placebo in a 6:2 ratio for 21 days. Subjects received the study medication in a once daily regimen after a light standardized breakfast. The participants in these panels were synchronized for their menstrual cycle; i.e. the initiation of dosing was always on Day3 ±2 of the menstrual cycle. The dose levels investigated were 20, 60 and 180 mg of compound k-5. Blood samples were collected on the day prior to dosing as well as on Day 21 (the final day of dosing) and plasma fractions were retained and stored frozen (−20° C.) until analysis. Leptin concentrations in these plasma samples were determined by ELISA assay (Human Leptin Kit: R&D Systems, Cat. N° DLP00) based upon linear regression analysis against a standard curve.

[0347] Effects on Leptin: FIG. 4 illustrates that compound k-5 treatment significantly (*p<0.05; paired t-test) increases serum leptin levels in women at all dose levels following 21 days of treatment relative to pre-dose levels. In comparison, placebo treatment had no significant effect on plasma leptin levels.

[0348] Study Conclusion: Compound k-5 increases circulating leptin levels in non-obese, premenopausal women. As leptin is a hormone known to have effects on weight gain and adiposity, these results may be relevant to %fat density relative to overall body composition, analogous to the data presented above for female, non-human primates (example 1).

Example 6: Effect on Plasma Leptin Level in Female Rats Over the Estrous Cycle

[0349] Rats do not have a menstrual cycle, but rather an estrous cycle that can be clearly defined by peak estrogen levels (precluding ovulation) that occurs regularly over a period of ˜4 days.

[0350] Daily plasma samples were collected each morning from sexually-mature female rats and analyzed for peak estrogen levels (coincident with proestrus) in order to define the timing of the estrous cycle for individual rats.

[0351] Next, rats were dosed orally BID (i.e. twice a day) with vehicle (0.5% methylcellulose) over a 4-day ‘baseline’ estrous cycle. Plasma samples were collected daily, each morning, and stored frozen prior to analysis for leptin.

[0352] Then, the rats were separated into two groups treated with either compound k-5 10 mg/kg, oral BID, or with vehicle over the 4-day ‘treatment’ estrous cycle. Plasma samples were collected daily, each morning, and stored frozen prior to analysis for leptin.

[0353] In a single experiment, plasma samples collected daily during both the ‘baseline’ and ‘treatment’ cycles were analyzed in a common experiment by ELISA assay based upon linear regression analysis against a standard curve. Leptin levels over the 4-day estrous cycle were calculated for individual rats by determination of AUC (area-under-the-curve) using credited data analysis software (GraphPad Prism).

[0354] Effects on Leptin: FIG. 5 illustrates that compound k-5 treatment significantly (*p<0.05; paired t-test) increases plasma leptin levels in female rats over the 4-day estrous cycle compared to when the same rats were treated with vehicle in the ‘baseline’ cycle.

[0355] Study Conclusion: Compound k-5 treatment increases circulating leptin levels in non-obese, sexually-mature female rats over the estrous cycle. As leptin is a hormone known to have effects on weight gain and adiposity, this finding may be relevant to % fat density relative to overall body composition, analogous to the data presented above for female, non-human primates (example 1) and in women (example 5).