Title of Invention	A PROCESS OF MAKING PLANT EXTRACT MIXTURES
Abstract	A composition comprising extract of one or more plants of one or more of the following plant families: Cissus, Vernonia and Brillantasia. Such compositions have beneficial activity principally in controlling weight gain and obesity, especially in conjunction with chitosan or chitosan derivative and an antioxidant such as vitamin C.

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FORM 2 THE PATENTS ACT, 1970 (39 Of 1970) COMPLETE SPECIFICATION See Section 10, rule 13] A PROCESS OF MAKING PLANT EXTRACT- MIXTURES MEDEX SCIENTIFIC (UK) LIMITED of THE OLD GRAIN STORE, 4 DENNE ROAD, HORSHAM, WEST SUSSEX RH12 1JE, a UNITED KINGDOM Company The following specification particularly describes the nature of the invention and the manner in which it is to be performed : - This invention is concerned with PROCESS OF MAKING PLANT EXTRACT MIXTURE from plants in at least one of three families (one or more of Cissus, Vernonia and Brillantasia), and uses of the mixtures to enhance fat-binding capacity as WELL as the inhibition of carbohydrate breakdown, amylase activity and nutrient absorbtion in the presence of fat binding materials. In particular the mixtures can be combined with chitosan or chitosan derivatives to synergise their fat binding properties. Fat binding materials e. g. chitosan have applications in industry and in health. In both cases, the binding capacity of these materials is limited because of their BULK. The new combination of plant extracts has the ability to enhance the fat binding capacity of such materials as WELL as alter the metabolism of other compounds in animals, including humans. Chitosan is widely used for the control of weight Its use is based on its ability to bind fatty acids in vitro and in vivo, thereby reducing the ability of the body to absorb and utilise dietary fats. By combining chitosan with two plant extracts, Cissus QUANDRAGULARIS and Veronia glabra, its ability to bind fatty acids and TR1ACYLGLYCEROLS in vitro was significantly increased (p 2 Chitosan is a polysaccharide produced from chilin found in Ihe Exoskeleon of arthropods (cruralcicoana and insecti) and the endoskolons of cophaloped : it is. widely spread in the biorriass, being the most abundant biopolymer after cellulose It is generally accepted that chitin is extensively acetylated while chitosan is largely deacetylated. Chitosan is a cationic" glucosamine polymer with a high anion-exchange capacity as a result of quaternary ammonium ions. It is known to have a marked hypocholesterolemic effect in rats, alters bile acid metabolism and increases HDHotal cholesterol ratio in broiler chickens. The hypocholesterolemic effect of chitosan can be theoretically explained by its ability to decrease lipid absorption and increase fecal cholesterol excretion. The alteration of bile acid can be due to the modification of colon pH. Chitosan is widely used in weight control products. Its application is a result of its ability to bind triglycerides in vitro, In principle therefore, chitosan limits the amount of fat energy that can be absorbed and used by the body. The use of chitosan therefore does not find application only as a weight loss supplement, but also as a means of reducing blood lipids. We have investigated the effect of chitosan and chitosan based formulations on overweight women. Their BMI and body composition had been evaluated. The fecal weight, fecal pH, water content, calcium and magnesium content and total nitrogen were measured. The total blood lipid content and beneficial effects of plant supplements (extracts of Vernonia glabra and Cissus quadranguians) on improved chitosan lipid binding has unexpectedly been found. plant families: Cissus. Verronia and Brillantasia According to another aspect the According to one aspect of the present invention there is provided a composition comprisina extract of one or more plants of one or more of the following invention provides uses of such compositions in treating disease or disorder, in particular for preventing, controlling or combatting obesity. In another aspect the invention -3- " provides such compositions for the preparation of a medicament for use in treating any of the ailmenls or conditions herein described. The composition preferably comprises or essentially consists of a mixture of at least two of the said plant extracts. The composition preferably also include one or more and bainading malenals such as chitosan or a chitonsan denvative, one or more arnylase inhibiting compounds and/or one or more antioxidants such as vitamins A, C, or E. The plant extracts are usefully obtained from dried or freeze dried plant components. The extract may be of the leaves, roots and/or stem of the plants. The extract may be dried powder or entirely aqueous-derived such extract, e.g. obtained by boiling the plant components in water, or derived from a mixture containing water and, for example, ethanol. A mixture of water and 50% by volume of ethanol is suitable and preferred. The extracts are also preferably ion exchange unbound and resin scavenged prior to use and formulation with chitosan or chitosan derivative, it is possible, but less preferred to use extracts derived from the solvent dichioromethane. preferably in the case of/plants of the Cissus family the plant components used in making the extract are the leaves, roots and/or stem. Preferably the extracts are aqueous and can beproduced by a simple, conventional extraction technique, although it is more preferable that the plant components be dried before addition of water and boiling the mixture. Such compositions can be formulated for human consumption and can demonstrate one or more of the following other properties: (1) Reduces the amount of fat absorbed by the body. (2) Increases the amount of fat in faeces. (3) Increases faecal bulk. (4) Reduces carbohydrate breakdown in vitro. - 4- (5) Reduces carbohydrate breakdown in vivo, (G) Inhibits salivary amylase activity, (7) Inhibits intestinal amylase activity, (8) Decreases the acidity of the stomach, (9) Increases the amount of cholesterol in faeces, (lO)Reduces post-prandial blood glucose, (11)inhibits intestinal lipase activity, (12)Reduces the body mass index (weight) of subjects. Example 1 Preparation of the mixture the Cissus family, Vernonia family, and Brillantasia family: Plant component used: Leaves, stems and roots Preparation method: The leaves, stems and roots are dried at 45°C for 72 hours. The dried samples are transferred into 2 times their weight of water and boiled for 1 hour. The mixture is ieft to stand for 2 hours at room temperature before straining to remove any residue. The resultant supernatant is stored at 40 C until required, Brillantasia sp. Plant component used: Leaves Preparation" method: The leaves are harvested from the plant and dried at 450 C for 72 hours. The dried leaves are finely ground into a powder, then transferred into 2 times their weight of boiling water. This is left to stand for two hours at room temperature before straining to remove leaf residue. The resultant supernatant was stored at 4o;C until required. Cissus sp, and Vernonia sp. combination Although all combinations of extracts of plants of the three_families._narnely Cissus sp. Vernonia sp. and Brillantasia sp. had an enhancing effect, optimal activity was obtained using the following mixture of two aqueous plant extracts: Cissus sp. extract 50-90%, more preferably 60-80% most preferably 70% (v/v) Vernonia sp. 10-50% more preferably 20-40% most preferably 30% (v/v) This is a preferred specific mixture of Cissus sp. and Vernonia sp. Application and Dose The mixture can be taken orally at a concentration of 0.1 to 10ml, preferably 0,1 to 5ml, more preferably 0.2 to 2ml, most preferably 0.5ml per kilogram body weight with or before a meal. Example 2 : Plant materialsVerponi glabra (root) andl Cissus quadrannguhms (leaf and slentI)were harvested from the Western and Centre Provinces of Cameroon respectively. The plant material was washed and dried for three days in an oven at 55oC. The dried material was then ground, seived and stored until required. Chitosan based formulations were made as shown in Table ] (increasing concentration of plant extract and decreasing chitosan content) Table 1 : Composition of different chitosan based formulations Constituents (mg) Chitosan formulations CF0 CF1 CF2 CF3 CF4 Chitosan 250 200 170 150 100 Vitamin C GO 60 60 60 60 Plant powder 0 50 80 100 150 Iiwitro binding studies The ability of various chitosan combinations to bind triacylglycerol was carried out as described below. A specific weight(l-4g) Of chitosan or the chitosan combination was weighed out and incubated with shaking at 37oC for 2 hours with 6g soya oil and 10 ml hydrochloric acid (10 mmol/L). The acid was then neutralised and 10 ml phosphate buffered saline (pH 7,4) added. Incubation with shaking was continued for a further 3 hours at 37°C, after which the tubes were centrifuged at 2500 rpm for 30 min. Unadsorbed fat floated on the surface, was aspirated and weighed. This gave the amount of triacylglycerol not bound to the chitosan combination, Subjects Seventy four overweight (BMI 25 - 30 kg/m2) women ( 19-32 years) were recruited to take part in.the study. They gave their written consent after details of the trial had been verbally explained to them and could drop out of the study at anytime they wanted without having to give reason. Ethics approval was obtained from the Joint University /Ministry of Health Ethics committee, Cameroon. Subjects were randomly allocated one of four treatments in a double blinded study. The results are for fifty nine subjects who completed the study. The control group received maize flour, while the other groups received either chitosan, chitosan plus Vemonia glabra (16% w/w) or chitosan plus Cissus quadrangulans (16% w/w). All subjects were required to consume 2g of the control or test material twice daily before their main meals. Diet Tlie subjects were asked not fo change their food habits and (o maintain [heir-normal diets as much as possible. They kept individual food diaries which were used to analyze food intake using food tables. Fecal collections and analysis . Faeces were collected for four consecutive days every other week, in special glass containers and brought daily to the laboratory for storage. All subjects were instructed to bring in their samples as soon as was possible. The feces was weighed and the pH measured. An aliquot (2 g) of the faeces was used for the determination of total lipid using the method described by Folch et al, (1957). The dry matter weight was determined using a homogenate of the total feces collected (3 dairying at 5 5°C). Fecal ash was determined after 48 hours incineration at 500°C. Fecal calcium and magnesium was determined using the modified methods described by SIGMA company. In these methods, the ash was dissolved in nitric acid solution (3N) and the calcium and magnesium assayed spectrophotometrically using arsenazo dye III and calmagite respectively. Total nitrogen was determined by the Kjeldahl procedure. Statistical significance was deiennined using paired Student"s t-tesi l In vitro lipid binding The in-vitro lipid binding of the different chitosan formulations is shown Table 2, The highest lipid binding was obtained with the formulation CFi for each of the two plant powders containing 16% (w/w) of the plant. Cissus quandragularis however bound more (P Table 2. The effect of Chitosan fonnularions on in-vitro lipid (Soja oil) binding (g/g of Chitosan formulation). mcamS.D Plant Powder Chitosan formulations CF0 CF, CF3 CFj CF, V.glabra 11.70*2.04 22.04*2.79** 17.81*4.09 12.49*4.78 10.44*4.9 C.quandragula I3.70±2.04 27.80*0.47** 22.60*6.52** 18.03*1.17* 14.89*2.05 *P **P Food intake The average energy and nutrient inlake of subjects is ijiven in Tnble i I hi:; , is similar lo the intake ofthal age group of women in Cameroon. Tabic 3: The mean of"total energy, lipid, carbohydrate, protein, calcium, magnesium and fiber of subject dietary intake mean ± SD ENERGY (Kcal/dav) 2346.48±535.55 LIPID (p/dav) 92.93±28.00 CARBOHYDRATE (p/dav) 367.51*64.54 PROTEIN (a/dflv) 83.50*15.15 FIBER (a/dav) 22.41±5.58 CALCIUM (mfi/dav) 42i.78±243.98 MAGNESIUM (mo/da v) 134.58±45.8G Fecal composition During the first week of the trial, only the formulation containing Vernonia glabra (16%, wAv) significantly (p The reduction of fecal pH and moisture content noticed is reversed by the inclusion of Cissus quadrangulatis. ^- This study has shown a negative correlation between the fecal moisture and the fecal pH (r-0.477) and even between fecal moisture and the root square of fecal pH [(pH) 1.2] (r-0.465), P The reduction of fecal moisture is also dependent on the ash content since fecal moisture is negatively correlated"to" the fecal ash content (mg/g of FDW) (r=-0.301), P Tablc 5; Correlation between different fecal components Variable r (N=80) Probabiliiy(P) ♦ moisture, pH -0.477 P ♦ moisture, (pH) " -0,465 P ♦ moisture, ash -0.301 PO.0I + ash ,pH +0,248 P ♦ calcium, pH -0.221 P ♦ moisture,calcium +0.030 N.S ♦ ash , calcium -0.069 N.S ♦ pH ,Nitrogen -0.232 PO.05 ♦ ash, Nitrogen +0.009 N.S ♦ moisture, Nitrogen +0.011 N.S Chitosan as well as the chitosan formulation containing Cissus quadrangulans significantly (p Fermentation of chitosan in the large intestine produces glucosamine which can be absorbed (application in arthritis) by the body. Glucosamine can also bring about an increase in gut pH which favors the absorption of nitrogenous compounds (negative correlation between fecal pH and total fecal nitrogen, r = -0.232; p Vemonia glabra therefore seems to play a role in inhibiting the absorption of nitrogenous compounds which will otherwise be favored by an increase in pH. /Chitosan as well as chitosan based formulations significantly (p Ho- Tablc 4: The effect of Chitosan formulations on fecal wet and dry weight, pH and moisture Measures Control (n=20) Chilnsan (n=21) Chitosan+V.g{n=18) Chilosan-K:.q(n=21) Do D>00 D0 D>00 Do D0 D0 D>00 Fecal wet weight(g/d) 165.4=26.4.a 187,8=41.5 a 185.7=24.6 a 180.2=16.7 a 178.5 =9.0a 264.2+33.3 188.1+28.6 a 187.8+82 a Feeal drv weigt(g/d ) 26.3=.5.3 a 28.5=9.8 a 29.0=5.3 a 28.9=9.7 a 28.2=8.2 a 36.0=94 b 28.2=7.7 a 27.2=9.4 a Feeal niciislure % 79.8=4.9 a 80.l=5.2 a 7S.6=5.9 a 72.6=6.7 b 79.1 =4.0 a 81.0=4.5 a 87.9=3.8 a 74.9=6.1 ab feeal pH 6.9=0.6 a 6.8=0.8 ac 6,9=0.4 a 7.4=0.3 b 6.8=0.2 a 6.5=0.3 c 6.9=0.5 a 7.0=0.3 a Vales are 4-days mcsnds= S.E Vale Without a common sepcrsenpl are significaotly dillerent (P Table 6; The effect of chitosan formulations on fecal ash, calcium and magnesium and nitrogen Measures Control (n=20) Chilosnae(n=2l) Chitosan+V.g(n=18) Chitisan+C.g(n=2n Do D,30 Do D>00 Do D>w D0 D,00 Feeal nsh g/d 2.5=1.1" 3.0=0.9 a 2.7=1.4 a 5.2=3.7 b 3.0=1.0 a 3.7=1.8 a 2.8=1.8 a 5.4=2.5 b Ciiteiiim mg/d 100.=23.7* 117.6=12.0 a 104.4=10 8 a 180.3=85.5 b 88.4=34.2 ac 65,1=15.2a 109.4=15.3 a 130.1=14.3 b Magnesium mp/d 83.5=21,3 a 76.5=16.9 a 89.6=17.6 a 100.5=32.5 a 82.3=11.4 a 89.4=14.2* 91.1*21.1" 92.5=l2.9a Nitrogen (N2) mg/d 354.5= 128.5* 384.3=194. 3" 39I.7=133. 6* 421.7=355. 0" 379.9=170. 9" 1039.5 =540 b 380.61.164. 5" 371.1=196. 4" N2-ChilosanN2 mg/d 33"15=l28.5* 384.3*194. 3" 39I.7=133. f.1 312.9=355. 379.9= 170. 9" 930.7-540. 1 380.6=164 5" 262.3. 196. 4C Values jrc d-d.iys meanstS.E -Slalixlical .significant;!: ri by cnmpnri.ion within a specific measured pnr.imtltr. V.tlnci willisnil .T coinnuiii n ipiuc script art; ,^1 pni[ii-.-unonly dibTt;;ni (p"0.0 5) Tablc 7: The effect of chitosan formulations on tola! lipid excretion Conirol (n=20) Chilos;inf (n=21) Chilos;an+V.g(n=18) Chitosan+C.q(n=2!) Do D,» Do D.m U D-00 Dn D 00 Tabla lipid (mg/g)"1 g/d 73±15a 87=15a 83 =l6a 163=18b 86=19* 183=12"" 88=I6* 161=6 b 8.4=3 7 l 10.0=4.0 9.6=4.1 a 29.4= 12 h 9.9=4.5" 48.3.)9.2b 10.213.9 a 30.3=13.2 a Values .ane:"l-day"s menus S.E Values-I itillitiul a ammiiis *spstcrptt ,and: "agnilic ane ilillera(l 0.01) Example 3 . The effect of chitosan based formulations on body composition in overweight women. Chitosan and chitosan based formulations containing Cissus quadrsngularis and Vernonia glabra were administered to sixteen overweight (BMI >25kg/m2) women on " an energy restricted diet supplemented with vitamins and minerals over a period of" eight months. There was a significant reduction in the BMI of subjects given chitosan as well as the chilosan based formulations (4 grams per day) containing the plant extracts (16%, w/w), with the most significant reduction observed in subjects who received the chitosan based formulation containing Cissus quadranguisris. A significant reduction in the circulating levels of cholesterol and total triglycerides was observed in all the chitosan based groups. Materials and Methods The following combinations were used for the study: Group 1. Maize flour -Control group Group 2. Chitosan plus vitamin C Group 3. Chitosan plus vitamin C plus Vemonia glabra Group 4. Chitosan plus vitamin C plus Cissus quadrangularis Subjects Thirty two overweight.(BMI > 25 kg/m2) women (24 - 36 years) were recruited for the study. They gave their written consent after details of the thai had been verbally explained to them. They could drop out of the trial at any time without need to explain their action. The results reported are for twenty subjects who completed the study. Subjects randomly allocated one of the four treatments above in a double blind study. All subjects were required to consume 2 g of the control or test material twice daily before their main meals. Diet The subjects were given a number of possible diets they could follow, which provided a tola! daily energy intake of 1500 kcal. Body mass" index (BMI) and Body fat content The BMI of subjects was measured using an electronic scale and a metre rule attached to the wall. The percentage body fat was determined by using bioeieclrical impedance measurements. Blood collection and sampling Venous blood (20 ml) was collected from the forearm of subjects, and ssrurn prepared from it was stored in 1ml vials at -70°C until required. The concentration of total cholesterol and triglycerides were determined using Sigma kits.... Results Body Mass Index fkq/m2). Values are means = sem. TABLE 8 ; Do D7 D15 D30 O60 D90 D12o D150 D180 D210 " Control 30.61 ± 2.03 31.61+ 2.03 29,41 + 2.42 29.21 ± 1.6B 29.03± 1.76 29.03+ 2.34 30.04± 1.67 29.87 + 1 .98 29.22± 2.10 28.69+-3.01 Chilosan 20.66+ 1.78 28.60+ 1,7B 27.31 + 2.02 26.02+ 2.01 26.03+ 1.67 25.34± 1.67 27.87+ 1.B7 28.02+1 .67 26.21= 2.02 25.04+ 1,53* Chilosan + Vernonia glabra 29,92+ 2.17 29.83+ 2.18 28.33+ 1.67 26.18+ 1.78 25.87+ 1.23* 25.34± 1.56" 25.02± 1.78* 24.98+1 .62* 25.38± 1.55* 24.88± 1.67* Chilosan + cissus quadrangularis 28,43+ 1.53 28,62i ■1.33 28.04+ 1.89 26.44+ 0.98 24.64+" 1.22" 23.40± 2.68" 23,55+ 2.67" 25.60=3 .56 24.36= 1.78* 24.31=. 1.50" *p Conclusions: Subjects on a daily average energy intake of approximately 1500 kcaL did not show any significant change in BMI. Subjects on the formulations containing Vernonia glabra as well as Cissus quadrangularis had reduced BMIs after being on the formulation for 60 days. -t-i- Body fat content (%) TABLE c ■ DQ D? Dt5 D30 DC0 | D60 I D120 D150 D160 D240 Contra) 39=9 41 + 12 38±14 37±12 39 ±8 37±6 38+8 36±6 38±6 36=6 (3715) [35±4) (33=3) (32=2) (31+3) Chilosan 37+10 38±8 38±6 35 + 9 (31=2) 34+7 34+6 32=3 35+8 33=6 33=5 (35*8) (33±2) (31±2) (29±2) Chilosan + Vernonia glabra 35i9 37±10 36±12 (32+3) 34=8 31+9 29+10 29±8 (24+3)1 (28±8) 28=8 28=9 j (35+.6J (30-2} (27+2) (25+3) Chilosan + cissus 37+6 37±8 36=9 (32+2) 33+6 29*8 24±9" 24±6" 24=7-* 23±4" 24=5 quadrangularis (35±4) (28+3) (25+3) (22±2) (21=2) *p (The figures in bracket represents values for subjects who were on a daily energy intake of 600 kcal rather than 1500 kcal). Conclusion: There is a significant decrease in total body fat in the groups treated with the chitosan formulations in subjects maintained on a diet of 1500 kcal per day. In subjects maintained on a 800 kcal per day diet, a decrease in body fat was observed in all groups. Total Blood Cholesterol (q/L) 1.19 ± 0.18* 0.85 ±0.09** 0.81 ± 0.09" 1.01 ±0.11** *p S0.01. Significant differences are by comparing to the control for Control Chitosan Chitosan + vernonia glabra Ch"ttosan + Cissus quadrangularis each time point. Day-0 1.70 ±0.12 1.60 ±0.06 2.07 ±0,11 1.94 ±0.14 *p Day 7 1.50 ±0.09 1.44 ±0.10 1.78 ±0.10 Day 15 1.54 0.18 0.73 ± 0.06" 1.42 ±0.08 Day 30 1.65 ± 0.15 0.92 ±0.05" 1.37 ± 0.11 day GO 1.58=0.11 0.84 ±0.12" 1.23 ±0.30* Conclusion: Chitosan and the chitosan formulation containing Cissus quadrangularis were the most effective in lowering blood cholesterol levels for any given time point. Considering the composition of the formulations containing the plant extracts, it is obvious that there is a component in Vernonie glabra that inhibits the reduction of blood cholesterol levels by chitosan. On the other hand, a component or components present in Cissus quadrangularis may potentate this reduction. Table Total Blood Triglycerides (q/L) Day 0 Day 7 Day 15 Day 30 day 60 Control 1.84*0.08 1.83 ±0.08 1.76 ± 0.10 1.89 ±0.09 1,76 ±0.18 Chitosan 1.26 ± 6.13 1.18 ±0.05 0.6S±0.14" 0,83 ±0.11* 0,74 ±0.10 Chitosan + 1.87 ±0.31 1.19 ±0.08* 0.82 ±0.09** 0.73 ±0.06" 0.63 ±0.08 vemonia glabra Chilosan + Cissus 1.84 ±0.32 1.49 ±0,27 0.62 ±0.14** 0.84 ±0.07-* 0.70 ±0,10 quadrangularis *p Conclusion: Chitosan and chitosan formulations significantly decreased the circulating concentrations of triglycerides. This is as a result of their ability to bind triglycerides in vitro as well as in vivo. The presence of plant extracts did not seem to have a potentiating effect on the ability of chitosan to bind triglycerides. The effect of Cissus quadrangularis and Vemonia glabra combination on blood lipid levels The mixture used in this part of the work had the following composition: Chitosan 61.8% Vitamin -C 19.0% ■■ Vemonia glabra .powder 5.7% Cissus quadrangularis powder 13.5% This mixture had a superior lipid binding capacity in vitro compared to other chitosan formulations, containing either Vemonia glabra or Cissus quadrangularis. In Vitro Lipid binding capacity of mixture 32.6 grams oleic acid per gram of mixture Effect of mixture on BMl in overweight adults (BMl > 25) Body Mass Index(k.g/m2). Values are means= sem. Conclusions: The chitosan formulation containing a mixture of Cissus quadrangularis and Vernonia glabra significantly reduced the BMi of overweight females faster than formulations containing either Cissus quadrangularis or Vernonia glabra. This synergistic effect is possibly as a result of a reduction in the available glucose substrate for use as a fuel (a-arnylase inhibition by Vernonia glabra). Fat ■ preferentially used as fuel. The preferred "specific mixture amongst within the scope of the invention, has application because of its effects upon different physiological processes. Preferred applications include the following: 1} Use in the management "of obesity, overweight, weight maintenance, slowing down the addition of weight, reduction of bounce back during weight loss and complications resulting from obesity. This includes conditions such as hyperlipidemia, hypercholesteremia and, inability to move freely. 2) Uses in the management of diabetes and complications resulting from diabetes. This includes hypergiycaemia, ketonuha and diabetic coma. 3} Use in controlling gastric acidosis. 4) Use in the relief of constipation. The above physiological processes are apparently brought about by action of the mixture on one or more of the following factors: Reduction of fat absorbed by the body; Increases in the amount of fat in faeces Increase of faecal bulk; Reduction of carbohydrate breakdown in vivo; Decrease in acidity of the stomach; Increase in the amount of cholesterol in the faeces; Reduction of post-prandiai blood glucose; and/or Reduction of Body mass index (BMI) and weight. Inhibition of salivary and pancreatic amylase activity Human salivary amylase (Sigma A 0521) and porcine pancreatic amylase (Sigma A3176) were used as starting material. The substrate used was starch and the formation of maltose was used to quantify and measure the activity of the amylase. One unit of activity of the I8 "mixture reduced the activity of salivary amylase by 50%, and (he activity of pancreatic amylase.by 65%. Decrease of acidity of the stomach Laboratory animals wore led diets containing the mixture (0.5ml/kg body weight) after an overnight fast. ■ The content of their stomachs had a higher pH than control animals. Humans who ingested the mixture produced faeces with a lower pH than control humans who had not ingested the mixture. Inhibition of pancreatic lipase activity Pancreatic lipase (Sigma.L9780) was used as starting material. One unit of the mixture is the quantity that will reduce the activity of 0.48 units of lipase by 50%. (One unit of lipase will liberate 1.0pi of 2-monogiycehde from 1,2-digtyceride per minute at 37°C,pH 8.1). We Claim : 1) A process of making plants extact mixtures of Cissus family, Vernonia family and Brillantasia family for wherein leaves, stem & roots are dried at 45*c for 72 hrs, said dried leaves stem & roots are finely ground into a powder & transferred into 2 times weight of their of boiling water for one hours; the said mixture is left to stand for 2 hours at room temperature before staining to remove leaf residue; the resultant supernatant stored at 4*c to obtain the plant extract mixture. 2) A process of making plants extact mixtures as claimed in claims 1 which essentially consists of said extract or extracts and chitosan or chitosan derivative. 3) A process of making plants extact mixtures as claimed in claims 1 further containing one or more amylase inhibiting compounds. 4) A process of making plants extact mixtures as claimed in claims 1, further containing one or more antioxidants. 5) A process of making plants extact mixtures as claimed in claim 1 in which the antioxidant is at LEAST one of Vitamin A, Vitamin C and Vitamin E. 6) A process of making plants extact mixtures as claimed in claim 1 in which the or each plant extract has been obtained by an aqueous based extraction process. 7) A process of making plants extact mixtures as claimed in claim 1 in which the or each plant extract has been obtained by an aqueous and ALCOHOL based extraction process. 8) A process of making plants extact mixtures as claimed in claim 1 wherein the or each extract is of one or more of the following plant families: Cissus 20 SP., BRILLANTASI sp. and Vernonia sp. and which preferably comprises a mixture of extracts derived from both Cissus sp. and Vernonia sp. 9) A process of making plants extact mixtures as claimed in claim 1 which comprises (on a volumetric basis) Cissus sp. extract 50-90% and Vernonia sp 10-50%. 10) A process of making plants extact mixtures as claimed in claim 1 in which the volume % of Cissus sp. extract is 60-80% and the volume % of Vernonia sp. extract is 20-40%. 11) A process of making plants extact mixtures as claimed in claim 1 in which the extract of plant of the Cissus family is extract of cissus quandragularis. 12) A process of making plants extact mixtures as claimed in claim 1 in which the extract of plant of the Vernonia family is extract of Vernonia GLABRA. 13) A process of making plants extact mixtures as claimed in claim 1 which ESSENTIALLY consists of extract of the plant Cissus QUANDRAGULARIS, chitosan, and optionally vitamin C. 21 Dated this 5th day of March, 2002.

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