FOOD SUPPLEMENT TO PROMOTE BODY WEIGHT LOSS

Abstract

The present invention describes a gastro-resistant pharmaceutical/nutraceutical composition comprising chincona cortex freeze-dried (Cinchona succirubra), chicory leaves and roots freeze-dried (Cichorium intybus, and/or Cichorium endivia, and/or Cichorium pumilum, and/or Cichorium spinosum), and gentian root freeze-dried (Gentiana lutea), and optionally griffonia seed extract (Griffonia simplicifolia). The nutraceutical composition object of the present invention is useful for appetite control, mealtime energy intake reduction, and body weight loss.

Claims

1. A gastro-resistant pharmaceutical/nutraceutical composition comprising: a. cinchona cortex freeze-dried; b. chicory leaves and roots freeze-dried; and c. gentian root freeze-dried.

2. The composition according to claim 1 further comprising Griffonia seeds extract.

3. The composition according to claim 1, wherein the cinchona is Cinchona succirubra and its cortex freeze-dried contains 1-3% by weight of quinine.

4. The composition according to claim 1, wherein the chicory is Cichorium intybus, and/or Cichorium endivia, and/or Cichorium pumilum, and/or Cichorium spinosum and its leaves and roots freeze-dried contains at most 350 mg/kg of lactucin.

5. The composition according to claim 1, wherein the gentian is Gentiana lutea and its root freeze-dried contains 1-2% by weight of gentiopicrin.

6. The composition according to claim 1, wherein the griffonia is Griffonia simplicifolia and its seed extract contains 98% by weight of 5-HTP.

7. The composition according to claim 1 comprising TABLE-US-00011 a. Chincona cortex freeze-dried 25-40%; b. Chicory leaves and roots freeze-dried 25-40%; c. gentian root freeze-dried 25-40%; where the percentages indicated are expressed by weight calculated with respect to the total weight of the composition.

8. The composition according to claim 1, wherein the three components are contained in a weight ratio of 1:1:1 and, when griffonia is present, the four components are contained in a weight ratio of 1:1:1:0.15.

9. The composition according to claim 1, formulated as gastro-resistant capsules or tablets.

10. A method of treatment of obesity and associated dysmetabolic diseases, said method comparing administering to a subject in need thereof a composition according to claim 1.

11. A method of appetite control and/or mealtime energy intake reduction and/or body weight loss said method comparing administering to a subject in need thereof a composition according to claim 1.

12. The composition according to claim 7, comprising TABLE-US-00012 a. Chincona cortex freeze-dried 30-35%; b. Chicory leaves and roots freeze-dried 30-35%; c. gentian root freeze-dried 30-35%; where the percentages indicated are expressed by weight calculated with respect to the total weight of the composition.

13. The composition according to claim 2, wherein the four components are contained in a weight ratio of 1:1:1:0.15.

14. The composition according to claim 2, comprising TABLE-US-00013 a. Chincona cortex freeze-dried 25-40%; b. Chicory leaves and roots freeze-dried 25-40%; c. gentian root freeze-dried 25-40%; d. griffonia seeds extract 2-15%; where the percentages indicated are expressed by weight calculated with respect to the total weight of the composition.

15. The composition according to claim 12, comprising TABLE-US-00014 a. Chincona cortex freeze-dried 30-35%; b. Chicory leaves and roots freeze-dried 30-35%; c. gentian root freeze-dried 30-35%; d. griffonia seeds extract 5-10%; where the percentages indicated are expressed by weight calculated with respect to the total weight of the composition

Description

DETAILED DESCRIPTION OF THE INVENTION

[0017] It was surprisingly found that the combination of these components produces a synergistic effect in decreasing body weight.

[0018] Preferably, the composition of the invention further comprises griffonia seeds extract (preferably of Griffonia simplicifolia).

[0019] Preferably, a composition according to the invention comprises:

TABLE-US-00001 Chincona cortex freeze-dried 25-40%; more preferably 30-35%; Chicory leaves and roots freeze-dried 25-40%; more preferably 30-35%; Gentian root freeze-dried 25-40%; more preferably 30-35%; [0020] where the percentages indicated are expressed by weight calculated with respect to the total weight of the composition.

[0021] In the composition according to the present invention, the three components, chincona cortex freeze-dried, chicory leaves and roots freeze-dried, and gentian root freeze-dried, are preferably contained in a weight ratio of 1:1:1.

[0022] Preferably, in accordance with a second embodiment, a composition according to the invention comprises:

TABLE-US-00002 Chincona cortex freeze-dried 25-40%; more preferably 30-35%; Chicory leaves and roots freeze-dried 25-40%; more preferably 30-35%; Gentian root freeze-dried 25-40%; more preferably 30-35%; Griffonia seeds extract 2-15%; more preferably 5-10%; [0023] where the percentages indicated are expressed by weight calculated with respect to the total weight of the composition.

[0024] In the second embodiment, of the composition according to the present invention, the four components, chincona cortex freeze-dried, chicory leaves and roots freeze-dried, gentian root freeze-dried, and griffonia seeds extract, are contained in a weight ratio of 1:1:1:0.15.

[0025] The composition will normally be formulated in gastro-resistant capsules or tablets, using known techniques (encapsulation, compression, controlled release, microgranules, nanocapsules, multilayer) as described in the pharmacopoeia for the preparation of pharmaceutical formulations for oral use.

[0026] A particular example of a pharmaceutical formulation according to the invention consists of gastro-resistant capsules containing: [0027] 100-300 mg of chincona cortex freeze-dried, preferably 200 mg; [0028] 100-300 mg of chicory leaves and roots freeze-dried, preferably 200 mg; [0029] 100-300 mg of gentian root freeze-dried, preferably 200 mg; [0030] optionally 10-50 mg of griffonia seeds extract, preferably 30 mg.

[0031] Particularly preferably, the composition may contain:

TABLE-US-00003 quinine 1-9 mg, preferably 6 mg; lactucin 0.35-1.05 mcg, preferably 0.70 mcg; gentiopicrin 2-6, preferably 4 mg; optionally 5-HTP 9.8-49 mg, preferably 29.4 mg.

[0032] A particular example of a pharmaceutical formulation according to the invention is a gastro-resistant capsule consisting of: [0033] chincona cortex freeze-dried, 200 mg; chicory leaves and roots freeze-dried, 200 mg; gentian root freeze-dried, 200 g. In particular, the composition preferably comprises quinine, 6 mg; lactucin, 0.70 mg; gentiopicrin, 4 mg; and optionally 5-HTP, 29.4 mg.

[0034] The composition according to the invention is obtained simply by mixing the components in the required amounts using normal mixers.

[0035] The present invention can be better understood in the light of the following embodiments.

EXPERIMENTAL PART

Example 1Preparation of Chincona Cortex Freeze-Dried

[0036] The chincona cortex was subjected to freezing at ?50 ?80? C., freeze-drying, and grinding.

TABLE-US-00004 Chincona cortex 1-3% quinine

Example 2Preparation of Chicory Leaves and Roots Freeze-Dried

[0037] The chicory leaves and roots were subjected to freezing at ?50 ?80? C., freeze-drying, and grinding.

TABLE-US-00005 Chicory leaves freeze-dried max 350 mg/kg lactucin

Example 3Preparation of Gentian Root Freeze-Dried

[0038] The gentian root was subjected to freezing at ?50 ?80? C., freeze-drying, and grinding.

TABLE-US-00006 Gentian root powder 1-2% gentiopicrin

Example 4Preparation of Griffonia Seeds Extract

[0039] Griffonia seeds were subjected to grinding and subsequent extraction with 95% ethyl alcohol. The alcoholic extract was separated by centrifugation and dried by spray-drying method.

TABLE-US-00007 Griffonia seeds dry extract 98% 5-HTP

Example 5Preparation of Gastro-Resistant Capsules

a. First Embodiment of the Invention (Mix1)

[0040] 800 mg gastro-resistant capsules, whose heads and bodies consists of 1.9% titanium dioxide, 5.0% gellan gum, hypromellose q.s. to 100%, were selected. A mixture consisting of freeze-dried and excipients (titanium dioxide 5 mg, corn starch 50 mg, microcrystalline cellulose 30 mg, calcium phosphate 5 mg, talc 5 mg, aluminium silicate 5 m), was prepared to fill the capsules so as to ensure an overall content as follows: chincona cortex freeze-dried, 200 mg; chicory leaves and roots freeze-dried, 200 mg; gentian root freeze-dried, 200 mg. In particular, the composition preferably comprises quinine, 6 mg; lactucin, 0.70 mg; gentiopicrin, 4 mg.

b. Second Embodiment of the Invention (Mix2)

[0041] 800 mg gastro-resistant capsules, whose heads and bodies consists of 1.9% titanium dioxide, 5.0% gellan gum, hypromellose q.s. to 100%, were selected. A mixture consisting of freeze-dried and excipients (titanium dioxide 5 mg, corn starch 50 mg, microcrystalline cellulose 30 mg, calcium phosphate 5 mg, talc 5 mg, aluminium silicate 5 m), in appropriate proportions, was prepared to fill the capsules so as to ensure an overall content as follows: chincona cortex freeze-dried, 200 mg; chicory leaves and roots freeze-dried, 200 mg; gentian root freeze-dried, 200 mg; griffonia seeds extract, 30 mg. In particular, the composition preferably comprises quinine, 6 mg; lactucin, 0.70 mg; gentiopicrin, 4 mg; 5-HTP, 29.4 mg.

Example 6Clinical Data

[0042] Study groups. 126 obese subjects, of both sexes, with a Body Mass Index (BMI)?30 kg/m.sup.2 in baseline conditions were recruited at the Ambulatorio del Programma Dipartimentale Terapia dietetica nei trapianti e nellinsufficienza renale cronica, Facolt? di Medicina Universit? Federico II, Naples. The subjects were randomized into 7 groups (18 patients each). Each group was treated with a low-calorie diet for 2 months, plus supplementation, as follows: Group 1, placebo, Group 2, chincona cortex freeze-dried; Group 3, gentian root freeze-dried; Group 4, chicory leaves and roots freeze-dried; Group 5, mix of the three freeze-drieds (mix1); Group 6, griffonia seeds extract; Group 7, mixture of the four components (mix2). Patients with diseases such as cancer, acute and chronic metabolic and inflammatory diseases, type 1 and type 2 diabetes treated with insulin and/or hypoglycemic drugs or treated with weight-loss drugs or hormone therapy were excluded. The study was approved by the Comitato Etico Istituzionale dellUniversit?degli Studi di Napoli Federico II; all patients gave their informed consent to the recruitment.

[0043] Dietary treatment. A personalized diet plan was set up for each patient of each group, with a 40% caloric restriction of the total energy requirement and the following nutrient composition, in accordance with the RNI (Recommended Nutrient Intake) guidelines: 55-60% of the total caloric intake from carbohydrates, 10-15% from proteins and 20-25% from fatty acids (<7% from saturated fat).

[0044] Supplementation. The subjects received a specific treatment, based on the group they belonged to, as follows: Group 1, 600 mg (two capsules)/die of placebo (maltodextrins); Group 2, 600 mg (two capsules)/die of chincona cortex freeze-dried; Group 3, 600 mg (two capsules)/die of gentian root freeze-dried; Group 4, 600 mg (two capsules)/die of chicory leaves and roots freeze-dried; Group 5, 600 mg (two capsules)/die of mixture (mix) of the three freeze-drieds (1:1:1); Group 6, 600 mg (two capsules)/die of griffonia seeds extract; Group 7, 630 mg (two capsules)/die of mixture (mix2) of the four components in a ratio of 1:1:1:0.15. The treatment was taken one hour before the main meals. Participants who did not take the treatment for two or more days were excluded from the study. Each patient received the right amount of treatment, provided by the NutraPharmaLabs Laboratories of the Dipartimento di Farmacia dellUniversit? degli Studi di Napoli Federico II, to be used for the entire duration of the study.

[0045] Study protocol. The subjects were evaluated at the time of recruitment (time TO), after 30 (time T1) and 60 (time T2) days of treatment, using standardized protocols. The nutritional status was assessed by anthropometric measurements: weight (Seca GmbH & Co KG, Hamburg, Germany), height (wall stadiometer with 0.1 cm accuracy), body mass index (BMI), waist circumference (WC), hip circumference (HC). To assess body composition, bioelectrical impedance analysis (BIA) was performed with a four-pole BI (RJL 101; Akern SRL, Florence, Italy). BIA was performed with a single frequency measurement (50 kHz). Visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) were evaluated in fasted subjects by the same operator with the Bodymetrix BX2000 instrument. In particular, VAT and SAT were measured 1 cm above the umbilicus at the end of exhalation and applying the same probe pressure for all subjects. Each measurement was performed 3 times and the average of the 3 measurements was used for the analysis. Hand muscle strength (kgm) was measured on dominant and non-dominant hands to the nearest kilogram, using a hand dynamometer (78010; Lafayette Instrument Company, Lafayette, IN, USA). During the measurement, the participant was standing, and the arm of the measured hand was unsupported and parallel to the body. Indirect calorimetry was performed in measuring energy expenditure to assess the energy needs, using the VMAX 2900 calorimeter. In particular, the calorimeter uses the amount of inspired and expired gas exchanges to calculate energy expenditure. The canopy system was used during the test with no compression mask in order to use a light and comfortable device without pressure or contact with the subject's face. The participant was placed in the supine position, free from physical and psychological stress, fasted and awake.

[0046] Compliance. Compliance with the dietary intervention was assessed with the aid of Food Frequency Questionnaires, monitoring food intake at baseline and every month, until the end of the study. Assessment of compliance with physical activity was verified by asking subjects to complete a physical activity questionnaire. Compliance with the supplements was assessed by means of a daily questionnaire that asked each volunteer how long the supplement was consumed, as well as assessing the presence of adverse events.

[0047] Statistical Analysis

[0048] Randomization

[0049] A total of 126 eligible patients (67 men and 59 women) were randomly assigned to 7 subgroups (18 subjects each). If a patient left the group before the intervention period, he would be replaced by the next eligible patient enrolled. Concealed allocation was performed by an Internet-based randomization program, stratified by study site. The list of random numbers was generated by an investigator with no clinical involvement in the study. Patients, doctors, laboratory technicians and trial staff (data analyst statisticians) were unaware of the treatment assignment.

[0050] Primary and Secondary Efficacy Outcomes

[0051] The primary endpoints measured were changes in body weight (BW) and body mass index (BMI). The secondary endpoints measured were changes in fat mass (FM) and lean mass (FFM). All considerations relating to untreated patient were blindly assessed at the main investigation site.

[0052] The decision-making process was performed based on a consensus document (unpublished standard operating procedure).

[0053] Safety

[0054] Safety was determined by adverse event reports, laboratory parameters regarding liver and kidney function, vital signs (blood pressure, pulse, height, weight, and body mass index), physical and neurological examinations. Safety was assessed for the entire treatment period on days 8, 35, 55 and 80, including adverse events occurring in the first three weeks after cessation of treatment.

[0055] Power-Analysis

[0056] For proportion analysis, the effect size is estimated from the relative risk RR frequencies between the two experimental groups (e.g., in the primary study RR corresponds to the probability of receiving a drug treatment/probability of not receiving it). Since there are no previous studies similar to this one available, the effect size estimates were conducted assuming the following scenarios: [0057] Risk in the placebo group: 99%, 90%, 80% [0058] Risk in the treatment groups: 70%, 50%, 30%

[0059] The resulting RR used to estimate the sample size are:

TABLE-US-00008 99% 90% 80% 70% 1.41 1.28 1.14 50% 1.98 1.80 1.60 30% 3.30 3.00 2.67

[0060] This indicates that the sample size estimates explore situations where the risk of receiving drug treatment in treated patients is from about 1.1 to 3.3 times lower than in the control.

[0061] The sample size calculations were performed with three 1-? power values equal to 0.80, 0.95 and 0.99, and significance level ?=0.05.

[0062] The following table shows the overall sample sizes for the various hypotheses formulated.

TABLE-US-00009 Risk in Risk in the the RR (1-?) = (1-?) = (1-?) = placebo treatment (placebo/ 0.99 0.95 0.80 group groups treatment) ? = 0.05 ? = 0.05 ? = 0.05 99% 30% 3.30 30 24 18 50% 1.98 56 44 30 70% 1.41 118 88 60 90% 30% 3.00 46 36 26 50% 1.80 96 72 48 70% 1.28 304 222 144 80% 30% 2.67 70 54 36 50% 1.60 190 126 90 70% 1.14 1408 1008 626

[0063] Based on the above, it is believed that an adequate number of patients could be equal to 126, in a study in which the following is assumed:

[0064] Statistics and Methodology

[0065] During the process, it became apparent that dropouts and incomplete diary documentation resulted in missing data that could not be adequately handled by the expected robust comparison. To manage the missing data structure, a negative binomial was used, i.e. a generalized linear mixed effects model (NB GLMM) which not only provides unbiased parameter estimates in case of missing random data, Missing at Random (MAR), but also provides reasonably stable results when the MAR intake is violated. Patients who did not provide any diary data (leading to zero evaluable days) were excluded from the MAR based primary efficacy analysis, according to an all observed data approach, as proposed by White and colleagues. This approach is statistically efficient without using multiple imputation techniques. Data retrieved after withdrawal of randomized study treatment were also included in the analysis. Unless otherwise indicated, all experimental results were expressed as the average?standard deviation (SD) of at least three replicates.

[0066] Statistical analysis of the data was performed by Student's t-test or two-way ANOVA, followed by Tukey-Kramer multiple comparison test to evaluate significant differences between a pair of media. Statistical heterogeneity was assessed using the Cochran test (p<0.1). The I2 statistic was also calculated, and I2>50% was considered as significant heterogeneity between studies. If significant heterogeneity between studies was demonstrated, a random-effects model was used. If not, the results were obtained from a fixed-effects model.

[0067] Percent change in the average and SD values was excluded when determining SD values for a result. If not available directly, SD values were calculated from standard errors, 95% CI, p-value, or t-value. Subgroup analyses, defined above, were performed to examine possible sources of heterogeneity within these studies and included health status, study design, type of intervention, duration, total dose, and Jadad score. Treatment effects were analyzed using PROC MIXED with treatment and period as fixed factors, subjects as random factors and baseline measurements as covariates and defined as weighted mean differences and 95% CI, and calculated for net changes in the evaluated parameters. Data that could not satisfy the criteria of homogeneity of variance (Levene's test) and normal distribution (as determined by the examination of the residual diagram and Shapiro-Wilks test) even after the logarithmic transformation, were analyzed by a non-parametric test (Friedman). The significance level (a value) was of 95% in all cases (P<0.05).

[0068] Analysis Set

[0069] The entire analysis set group included all randomized patients and patients who did not fail to meet any important entry criteria. Patients who did not provide primary efficacy data from efficacy analyses were excluded. The protocol set consisted of all patients who did not substantially deviate from the protocol and they exhibited two characteristics: first, this group included patients for whom no major protocol violations were detected (e.g., poor compliance, errors in the assignment of treatment); second, they had to be treated for at least 50 days, starting on the day of the first intake (completion of a certain pre-specified minimum exposure to the treatment regimen). Therefore, patients who discontinued the study or treatment prematurely were excluded from the per-protocol sample.

[0070] Results

[0071] As can be seen from the data shown in Table 1, the formulation named mix1, consisting in the treatment of subjects belonging to Group 5, and comprising the mixture (mix1) of the three freeze-drieds in equal ratios (1:1:1), favored a decrease in body weight (BW), body mass index (BMI), fat mass (FM), and an increase in lean mass (FFM), greater than the sum of the effects of the components taken individually.

[0072] This formulation clearly indicates a synergistic effect of the various constituents of the composition.

[0073] The formulation named mix2, consisting in the treatment of subjects belonging to Group 7, and including the mixture (mix2) of the four components in a ratio of (1:1:1:0.15) led to results similar to those observed for mix1. The addition of griffonia seeds extract in mix2 therefore does not alter the synergistic effect observed for mix1.

CONCLUSIONS

[0074] The nutraceutical product based on a mixture of cinchona cortex freeze-dried, gentian root freeze-dried and chicory leaves and roots freeze-dried, in equal ratios (1:1:1), or based on a mixture of cinchona cortex freeze-dried, gentian root freeze-dried and chicory leaves and roots freeze-dried, and griffonia seeds extract in a ratio of (1:1:1:0.15), is provided as a useful support, or even a possible alternative, to classical pharmacotherapy for body weight loss and body composition control. The results obtained indicate the possibility of an innovative treatment that may represent a valid alternative for the treatment of obesity and associated metabolic diseases in the clinical practice.

TABLE-US-00010 TABLE 1 Parameters BW (kg) BMI (kg/m.sup.2) FM (%) FFM (%) Group 1 T0 97.8 ? 1.2 36.7 ? 0.4 41.5 ? 1.0 58.5 ? 0.9 (placebo) T1 93.0 ? 0.9 34.9 ? 0.3 39.7 ? 0.4 60.3 ? 0.3 ?% ?4.9 ?4.9 ?4.3 +3.1 Group 2 T0 94.8 ? 1.1 38.2 ? 1.0 37.9 ? 0.9 62.1 ? 0.7 (chincona) T1 79.8 ? 1.2 32.1 ? 1.0 27.9 ? 1.7 72.1 ? 1.0 ?% ?15.8 ?15.7 ?26.5 +16.2 Group 3 T0 102.8 ? 1.4 37.9 ? 0.3 44.1 ? 0.6 56.2 ? 0.5 (gentian) T1 92.0 ? 1.5 34.0 ? 0.4 35.1 ? 0.6 62.7 ? 0.5 ?% ?10.5 ?10.2 ?20.2 +11.6 Group 4 T0 98.1 ? 1.4 38.9 ? 0.3 39.3 ? 0.6 60.1 ? 0.5 (chicory) T1 91.0 ? 1.2 35.9 ? 0.4 33.3 ? 1.0 64.4 ? 0.9 ?% ?7.2 ?7.6 ?15.3 +7.2 Group 5 T0 100.1 ? 1.2 37.4 ? 1.0 40.4 ? 1.7 57.5 ? 1.0 (mix1) T1 84.3 ? 1.1 31.5 ? 1.0 30.0 ? 0.9 67.8 ? 0.7 ?% ?18 ?17.9 ?28.2 +17.9 Group 6 T0 97.8 ? 1.3 37.5 ? 0.4 38.2 ? 0.5 61.7 ? 0.4 (griffonia) T1 91.9 ? 1.1 35.3 ? 0.3 34.1 ? 1.1 65.8 ? 0.7 ?% ?6.0 ?5.9 ?10.7 +6.5 Group 7 T0 98.9 ? 1.3 38.3 ? 1.1 39.2 ? 1.5 58.3 ? 1.1 (mix2) T1 81.8 ? 1.2 31.4 ? 1.0 28.0 ? 0.7 68.5 ? 0.7 ?% ?17.3 ?18.0 ?28.5 +17.5 Group 1: treatment with placebo; Group 2: treatment with chincona cortex freeze-dried; Group 3: treatment with gentian root freeze-dried; Group 4: treatment with chicory leaves and roots freeze-dried; Group 5: treatment with the mixture of three freeze-drieds; Group 6: treatment with griffonia seeds extract; Group 7: treatment with the mixture of the four samples; T0, baseline state; T2, after 60 days of treatment; BW, body weight (kg); BMI, body mass index (kg/m.sup.2); FM, fat mass (%); FFM, lean mass (%). Data are expressed as average ? standard deviation. *p < 0.05