ORAL SUSPENSION WITH ANTIULCEROUS AND CHEMOPROTECTIVE EFFECT ON COLON CANCER AND PREPARATION METHOD THEREOF

Abstract

A new pharmaceutical composition consisting of an oral aqueous suspension for the prevention and treatment of gastric ulcers and colon cancer is disclosed, as well as the procedure for preparing it. This suspension is characterized by containing a purified extract with fatty alcohols from beeswax, microcrystalline cellulose and excipients accepted by the pharmaceutical industry. The method of preparing this suspension is characterized in that the fatty alcohol extract is subjected to a process of particle size reduction to <1.5 micrometers in the presence of an emulsifying agent at >1000 rpm; the mixture of said active ingredient with microcrystalline cellulose in the proportion and concentration in which they are formulated, makes these components act synergistically, conferring powerful antiulcer and chemoprotective effects to this composition.

Claims

1. Oral suspension with antiulcer and chemoprotective effect on colon cancer, characterized by being an aqueous suspension containing as active ingredients a purified extract of fatty alcohols from the refined wax of Apis mellifera bees and microcrystalline cellulose.

2. Oral suspension according to claim 1, characterized in that the ingredients are in the following proportion: 1-2% of the fatty alcohol extract; 1-2% microcrystalline cellulose; 2-4% emulsifying agents; 0.1-0.6% preserving agents; 20-30% moisturizers; 1-6% sweeteners; 0.1-1% suspending agents; 0.01-0.05% flavorings and 50-70% purified water.

3. Oral suspension according to claim 2, characterized in that the particle size of the purified extract of fatty alcohols from the refined beeswax is less than 1.5 micrometers.

4. Oral suspension according to claim 2, characterized in that the emulsifying agent is polyoxyethylene hydrogenated castor oil or polysorbate, the preserving agents are methyl and propylparabens, the wetting agent is glycerin, the sweeteners are sorbitol or sodium saccharin, the suspending agent is carboxymethylcellulose and the flavoring is an essential oil.

5. Procedure for obtaining an aqueous suspension with an antiulcer and chemoprotective effect on colon cancer, characterized by: A) mixing a purified extract of fatty alcohols with one or more emulsifying agents; B) subjecting said mixture to a technological process to reduce the particle size of the extract to <1.5 micrometers; C) incorporating into the mixture with constant stirring at >1000 rpm microcrystalline cellulose and preservative, wetting, sweetening, suspending, flavoring agents and purified water.

6. Process for obtaining an aqueous suspension, according to claim 5, characterized in that the fatty alcohol extract is extracted and purified from Apis mellifera beeswax.

7. Procedure for obtaining an aqueous suspension, according to claim 5, characterized in that the technological process applied to reduce the particle size of the fatty alcohol extract is based on subjecting the mixture of said extract and the emulsifying agent to a shearing process with a homogenizing equipment at a stirring speed >1000 rpm, at a temperature >80° C. for >1 minute.

8. Procedure for obtaining an aqueous suspension, according to claim 5, characterized in that the technological process applied to reduce the particle size of the fatty alcohol extract is based on subjecting the mixture of said extract and the emulsifying agent to a stirring speed >1000 rpm at a temperature >80° C. for >1 minute and then reduce the temperature to <60° C. with the same stirring speed.

Description

EXAMPLE 1

[0013] In an auxiliary stainless-steel container with a 50 L capacity (R1), 12 L of purified water were added and 1 kg of microcrystalline cellulose was incorporated little by little, at intervals and with stirring. It was stirred at 60 rpm for 30 minutes, until a homogeneous paste was obtained. 5 kg of 70% sorbitol and 24 kg of glycerin were added with stirring and stirring was continued for 10 minutes to obtain Mixture 1 (M1). On the other hand, in a stainless-steel reactor with a 300 L capacity, 60 L of water were heated to 90° C., 10 liters were separated in an auxiliary container (R2), 3 kg of polysorbate 80 were incorporated with stirring at 1200 rpm and it was stirred for 5 minutes at 1200 rpm. 1.2 kg of the purified extract of fatty alcohols was added little by little and at intervals and stirring was maintained for 10 minutes to obtain Mixture 2 (M2), after which the temperature was lowered to 30±2° C., maintaining the agitation. In another vessel, 0.18 kg of methylparaben and 0.02 kg of propylparaben were dissolved in 1.5 liters of ethanol with stirring at 60 rpm. 0.2 kg of sodium carboxymethylcellulose A/V were moistened, 0.03 kg of flavoring were incorporated and stirred for 5 minutes to obtain Mixture 3 (M3).

[0014] In another 5 L auxiliary vessel, 1 kg of glycerin was mixed with 0.5 L of ethanol and stirred for 5 minutes at 60 rpm to obtain Mixture 4 (M4). In the reactor where the mixture M2 was found, M1 was incorporated, dragging the remains with the minimum amount of water separated in R2 and stirred for 10 minutes at 60 rpm. After this time, M3 was incorporated and the remains were dragged with M4. Subsequently, what remained of the suspension was dragged with the hot water that was in R2 and stirred for 10 minutes. The stirring was then stopped and the mixture was allowed to stand for 10 hours. After this time, the remaining alcohol was sprinkled on the surface of the suspension and made up to the mark with water to complete 100 liters of suspension. It was stirred for 5 minutes at 1200 rpm, the suspension was filtered and a sample was taken for quality control. The final preparation was packed in amber glass bottles that were covered with plastic screw caps and the bottles were stored at room temperature.

EXAMPLE 2

[0015] In an auxiliary stainless-steel container with a 50 L capacity (R1), 12 L of purified water were added and 1 kg of microcrystalline cellulose was incorporated little by little, at intervals and with stirring at 40 rpm. It was stirred for 30 minutes, until a homogeneous paste was obtained. 5 kg of 70% sorbitol and 24 kg of glycerin were added with stirring and stirred at 40 rpm for 10 minutes to obtain Mixture 1 (M1). On the other hand, in a stainless-steel reactor with a 300 L capacity, equipped with a Polytron-type homogenizer system, 60 L of water were heated to 90° C., 10 liters were separated in an auxiliary container (R2), and incorporated with stirring. 9000 rpm 3 kg of cremophor RH 40 and stirred for 5 minutes. 1.2 kg of the purified extract of fatty alcohols was added little by little and at intervals and stirring was maintained for 10 minutes to obtain Mixture 2 (M2), after which the temperature was lowered to 30±2° C., maintaining the agitation. In another container, 0.18 kg of methylparaben and 0.02 kg of propylparaben were dissolved in 1.5 liters of ethanol with stirring at 40 rpm. 0.3 kg of sodium carboxymethylcellulose A/V were moistened, 0.03 kg of flavoring were incorporated and stirred for 5 minutes to obtain Mixture 3 (M3).

[0016] In another 5 L auxiliary vessel, 1 kg of glycerin was mixed with 0.5 L of ethanol and stirred for 5 minutes to obtain Mixture 4 (M4). In the reactor where the mixture M2 was found, M1 was incorporated, dragging the remains with the minimum amount of water separated in R2 and stirred for 10 minutes at 40 rpm. After this time, M3 was incorporated and the remains were dragged with M4. Subsequently, what remained of the suspension was dragged with the hot water that was in R2 and stirred for 10 minutes at 1000 rpm. The stirring was then stopped and the mixture was allowed to stand for 12 hours. After this time, the remaining alcohol was sprinkled on the surface of the suspension and made up to the mark with water to complete 100 liters of suspension. It was stirred for 5 minutes at 1000 rpm, the suspension was filtered and a sample was taken for quality control. The final preparation was packed in amber glass bottles that were covered with plastic screw caps and the bottles were stored at room temperature.

EXAMPLE 3

[0017] The composition obtained in example 1 (FAMC) was subjected to a preclinical test with animals, in which its antiulcer effect was compared with the antiulcer effects of the registered fatty alcohol tablet (RAT), and suspensions of Fatty Alcohols (FA) and microcrystalline cellulose (MC), respectively, the latter two prepared with doses of FA and MC equal to the doses of both substances in the FAMC suspension. The experimental model used in this trial was the induction of gastric ulcers in rats by sodium hydroxide.

[0018] Male Sprague-Dawley rats (250-300 g) were used, which were adapted for 7 days to the usual laboratory conditions at 20-25° C., relative humidity of 60±5%, light/dark cycles of 12 hours and with free access to water and standard feed for rodents. After completing the quarantine, the animals were randomly distributed into 10 experimental groups (10 rats/group): a negative control that only received the vehicle and 9 groups in which the ulcer was induced with 0.2 N sodium hydroxide, among these a positive control not treated with any antiulcer agent and 8 groups treated with the substances to be evaluated as shown in Table 1. The suspensions were administered as they are, while the RAT was prepared in acacia gum/water vehicle (1%). The use of a negative control group (not subjected to damage with sodium hydroxide), as well as that of a positive control group (subjected to damage and without treatment with any substance) allowed corroborating the validity of the model under the experimental conditions, and comparing the anti-ulcer efficacy of the substances evaluated.

[0019] All treatments and sodium hydroxide were administered orally, as a single dose, by intragastric intubation. The selected doses are within the range of effective doses proven in previous preliminary experiences. To induce gastric ulcer, the animals were fasted for 24 hours prior to the experiment, with free access to water. One hour after the administration of single doses of the vehicle, and the different treatments, each rat received the ulcer-inducing agent (1 mL/200 g) by gastric intubation. One hour later, the rats were sacrificed in a halothane atmosphere, the stomachs were removed and opened at the greater curvature.

[0020] Gastric mucosal lesions were quantified by two independent blinded observers. Lesion score was defined as the sum total of lesion sizes in mm.sup.2 (Ohara 1992). Statistical comparison between the mean ulcer areas in the different groups was performed using the non-parametric Mann Whitney U test. A priori, a significance level of α=0.05 was set and the commercial package Statistic for Windows (Release 4.2, StatSoft, Inc USA) was used.

TABLE-US-00001 TABLE 1 Effects of FAMC, MC, FA suspensions, and RAT on sodium hydroxide-induced gastric ulcer in rats. Doses Ulcer index Treatment (mg/kg/day) (mm2) I (%) Negative control 0    0 ± 0.00*** — (Vehicle) Positive Control 0 83.70 ± 13.8 — (Vehicle + NaOH) MC + NaOH 25   80 ± 10.6 4.4 MC + NaOH 200  75.4 ± 12.5 9.9 FA + NaOH 25  67.2 ±11.2 19.7 FA + NaOH 200  50.3 ± 13.5** 39.9 RAT + NaOH 25  51.4 ± 10.4 ** Δ 38.6 RAT + NaOH 200  40.5 ± 12.70*** Δ 51.6 FAMC + NaOH 25  11.3 ± 3.00*** °ab 86.5 FAMC + NaOH 200   12 ± 0.3*** °ab 98.6 FAMC: Suspension of fatty alcohols plus microcrystalline cellulose MC: Microcrystalline cellulose suspension FA: Suspension of Fatty Alcohols RAT: Fatty alcohol tablet registered as a nutritional supplement I inhibition, Data as Mean ± SEM (standard error of the mean) * p < 0.05, ** p < 0.01, *** p < 0.0001, comparison vs positive control Δ p < 0.05, °p < 0.0001 comparison vs similar dose of FA, a p < 0.0001 comparison vs similar doses of MC b p < 0.0001 comparison vs similar doses of RAT

[0021] As shown in Table 1, the oral administration of NaOH (0.2 N) produced gastric lesions in the animals of the positive control group compared to the healthy animals (negative control), which demonstrates the validity of the model in the experimental conditions

[0022] Oral administration with FAMC, FA, and RAT reduced the gastric ulcer index, while CM only produced slight reductions that were not significant. The FAMC suspension (25 and 200 mg/kg) produced a reduction in the gastric ulcer index, which was significant not only compared to the positive control group, but also compared to the same doses of the MC, FA and RAT suspensions. Therefore, the highest anti-ulcer efficacy was achieved with the FAMC suspension, especially since it produced ˜100% (total) inhibition of the gastric ulcer index with the highest dose tested of 200 mg/kg.

[0023] In addition, the FAMC suspension produced higher inhibition percentages than the sum of those achieved with the MC and FA suspensions separately, showing a synergism in the FAMC, where the presence of the FA extract and the MC in the proportions and concentrations in which appear promotes greater anti-ulcer efficacy.

[0024] It should also be noted that the antiulcer effect of the FAMC suspension is much higher than the effect reported for the 12% CM suspension (Barzaga 2004), both evaluated in alkaline gastritis ulcer induction models. Thus, the percentages of inhibition achieved with 25 and 200 mg/kg of the FAMC suspension (86.5 and 98.6%, respectively) are higher than that reported for MC, which was 50% at a higher dose (356.38 mg/kg) (Barzaga 2004), which is in correspondence with the synergistic effect of the FAMC observed in this example.

[0025] It is also worth noting that, in the present example, the lowest dose tested of 25 mg/kg of the FAMC suspension produced a high percentage of inhibition of 86.5% on gastric ulcers, which indicates that in addition to a high efficacy, the FAMC suspension also has high power.

EXAMPLE 4

[0026] The composition obtained in example 2 (FAMC) was subjected to a preclinical test with animals, in which its antiulcer effect was compared with the antiulcer effects of the registered fatty alcohol tablet (RAT), and suspensions of Fatty Alcohols (FA) and microcrystalline cellulose (MC), the latter two prepared with doses of FA and MC equal to the doses of both substances in the FAMC suspension. The experimental model used in this trial was the induction of gastric ulcers in rats by ethanol.

[0027] Male Sprague-Dawley rats (250-300 g) were used, which were adapted for 7 days to the usual laboratory conditions at 20-25° C., relative humidity of 60±5%, light/dark cycles of 12 hours and with free access to water and standard feed for rodents. After completing the quarantine, the animals were randomly distributed into 11 experimental groups (10 rats/group): a negative control that only received the vehicle and 10 groups in which the ulcer was induced with 60% ethanol, among these a control positive not treated with any antiulcer agent, 8 groups treated with the substances to be evaluated and a group treated with Omeprazole as a reference substance, as shown in Table 2. The suspensions were administered as they are, while a tablet was prepared in a suspension with 1% acacia gum.

[0028] The use of a negative control group (not subjected to damage with ethanol), as well as a positive control group (subjected to damage and without treatment with any substance) made it possible to verify the functioning of the model and compare the anti-ulcer efficacy of the substances evaluated. All treatments and ethanol were administered orally, as a single dose, by intragastric intubation. The selected doses are within the range of effective doses verified in previous preliminary experiences and the dose of omeprazole (20 mg/kg) has also been effective in this model. To induce gastric ulcer, the animals were fasted for 24 hours prior to the experiment, with free access to water. One hour after the administration of single doses of the vehicle, and the different treatments, each rat received the ulcer-inducing agent (1 mL/200 g) by gastric intubation. One hour later the rats were sacrificed in a halothane atmosphere, the stomachs were removed and opened at the greater curvature. The quantification of the gastric ulcer index was performed as described in example 3.

[0029] The results on the ethanol-induced ulcer index (table 2) show that the oral administration of ethanol produced the formation of gastric lesions in the animals of the positive control group compared to the animals of the negative control group (healthy). The animals treated with the FAMC suspension at doses of 25 and 200 mg/kg were the ones that showed a lower ulcer index compared to the animals with damage (positive control). Treatment with said suspension markedly and significantly decreased the gastric ulcer index, reaching 93.95 and 99.85% inhibition at doses of 25 and 200 mg/kg, respectively. Statistical comparisons between the effects of the different treatments used, at similar doses, showed significant differences, highlighting that the FAMC suspension showed the best results, even higher than the sum of the percentages of inhibition of the FA and MC suspensions, demonstrating a greater protection of the gastric mucosa. The reference substance used (Omeprazole) also decreased the gastric ulcer index, which corroborates the validity of the results obtained in our experimental conditions. It is noteworthy that the FAMC suspension achieved superior effects to those of Omeprazole, although they are not administered at the same doses, since it was not the objective of the study to compare both substances.

TABLE-US-00002 TABLE 2 Effects of FAMC, MC, FA suspensions, and RAT on ethanol-induced gastric ulcer in rats. Doses Ulcer Index Treatment (mg/kg/day) (mm.sup.2) I (%) Negative Control 0  0.15 ± 0.07*** — (Vehicle) Positive Control 0 169.09 ± 12.8 — (Vehicle) MC+ ethanol 25  155.5 ± 11.6 8 MC+ ethanol 200  133.3 ± 13.7 21.1 FA + ethanol 25 149.13 ± 12.2 11.7 FA + ethanol 200  89.19 ± 14.5** 47.3 RAT + ethanol 25 106.60 ± 11.40** Δ 37 RAT + ethanol 200  41.74 ± 13.70*** Δ 75.4 OMP + ethanol 20  35.97 ± 14.48*** 78.8 FAMC + ethanol 25  10.38 ± 3.00*** °a b 93.95 FAMC + ethanol 200  0.40 ± 0.34*** °a b 99.85 FAMC: Suspension of fatty alcohols plus microcrystalline cellulose MC: Microcrystalline cellulose suspension FA: Suspension of Fatty Alcohols RAT: Fatty alcohol tablet registered as a nutritional supplement I inhibition, Data as Mean ± SEM (standard error of the mean) * p < 0.05, ** p < 0.01, *** p < 0.0001, comparison vs positive control Δ p < 0.05, ° p < 0.0001 comparison vs similar dose of FA, a p < 0.0001 comparison vs similar doses of MC b p < 0.0001 comparison vs similar doses of RAT(Mann Whitney U test)

[0030] As a conclusion of the test presented in this example, it can be stated that oral administration with the FAMC suspension markedly and significantly inhibited the formation of gastric ulcer induced by ethanol in rats, with a higher efficacy than that achieved with the registered tablet and with the suspensions FA and MC at the same doses. Additionally, it was observed that the efficacy of the FAMC suspension was greater than the sum of the efficacy of the FA and MC suspensions in both doses evaluated, which demonstrates the existence of a synergistic effect in the composition object of the present invention.

EXAMPLE 5

[0031] The composition obtained in example 2 (FAMC) was subjected to a preclinical test with animals, in which its antiulcer effect was compared with the antiulcer effects of the registered fatty alcohol tablet (RAT), and suspensions of Fatty Alcohols (FA) and microcrystalline cellulose (MC), the latter two prepared with doses of FA and MC equal to the doses of both substances in the FAMC suspension. The experimental model used in this test was that of the induction of gastric ulcers in rats by ligation of the pylorus, which is an acid-dependent model, unlike the model presented in the previous example.

[0032] Male Sprague-Dawley rats (250-300 g) were used, which were adapted for 7 days to the usual laboratory conditions at 20-25° C., relative humidity of 60±5%, light/dark cycles of 12 hours and with free access to water and standard feed for rodents. After completing the quarantine, the animals were randomly distributed into 11 experimental groups (10 rats/group): a negative control that only received the vehicle and 10 groups in which the ulcer was induced by ligation of the pylorus, among these a positive control not treated with any antiulcer agent, 8 groups treated with the substances to be evaluated and a group treated with Omeprazole as a reference substance, as shown in Table 3. The suspensions were administered as they are, while a suspension with 1% acacia gum was prepared to administer the tablet. The use of a negative control group (not subjected to pylorus ligation), as well as that of a positive control (subjected to damage and without treatment with any substance) made it possible to verify the functioning of the model and compare the anti-ulcer efficacy of the substances evaluated.

[0033] All treatments were administered orally, as a single dose, by intragastric intubation. The selected doses are within the range of effective doses verified in previous preliminary experiences and the dose of omeprazole (20 mg/kg) has also been effective in this model. To induce gastric ulcer, the animals were fasted for 24 hours prior to the experiment, with free access to water. One hour after the administration of single doses of the vehicle, and the different treatments, the animals were anesthetized by intraperitoneal injection with sodium thiopental (35 mg/kg), a 2 cm-long incision was made in the anterior third of the line abdominal media, locating the stomach and ligating the pyloric sphincter with silk suture number 2 and the abdominal wall was sutured with silk thread. One hour later, the rats were sacrificed in a halothane atmosphere, the stomachs were removed and the gastric juice collected in test tubes. The latter was centrifuged at 3000 rpm, the supernatant separated and quantified in ml. The stomachs were opened through the greater curvature and the gastric ulcer index was quantified as described in example 3.

[0034] The results on the index of ulcer induced by pylorus ligation (Table 3) show that the formation of gastric lesions occurred in the animals of the positive control group with respect to the animals of the negative control group (healthy). The animals treated with the FAMC suspension at doses of 25 and 200 mg/kg were the ones that showed a lower ulcer index compared to the animals with damage (positive control). Treatment with said suspension markedly and significantly decreased the gastric ulcer index, reaching 76.3 and 99.7% inhibition at doses of 25 and 200 mg/kg, respectively. Statistical comparisons between the effects of the different treatments at similar doses showed that the FAMC suspension is the substance evaluated that presents the greatest anti-ulcer efficacy, since it was significantly superior to RAT, and to the suspensions with the respective monotherapies of FA and MC, respectively. In addition, the fact that the inhibition achieved by the FAMC suspension was greater than the sum of those achieved individually with MC and FA, demonstrates the synergistic effect of the presence of these two components in the mixture of the FAMC suspension.

[0035] Meanwhile, Omeprazole, the reference substance, significantly decreased the gastric ulcer index, which corroborates the validity of the results obtained in our experimental conditions. The volume of gastric juice was not modified by the suspensions of FAMC, MC, FA, or by RAT, which suggests that the antiulcer action of these substances does not depend on an effect on the formation of gastric acid. Only omeprazole reduced this variable, which is consistent with its mechanism of action as an antisecretory proton pump inhibitor.

TABLE-US-00003 TABLE 3 Effects of FAMC, MC, FA suspensions, and RAT on gastric ulcer induced by pylorus ligation in rats. Ulcer Index Gastric juice Treatment (Doses mg/kg/day) (mm.sup.2) I (%) volume Negative Control (Vehicle) 0  0,0 ± 0,00*** — 0,0 ± 0,00*** Positive Control (Vehicle + 0 23,2 ± 3,2 — 9,9 ± 1,2 pylorus ligation) MC + pylorus ligation 25 21,1 ± 1,6 9  10 ± 1,05 MC + pylorus ligation 200 20,5 ± 1,7 11,6 9,5 ± 1,2 FA + pylorus ligation 25 18,3 ± 2,2 21,1 9,8 ± 1,3 FA + pylorus ligation 200 13,3 ± 3,1** 42,7 9,7 ±1,1 RAT + pylorus ligation 25 16,1 ± 2,40** 30,6 9,8 ±1,1 RAT + pylorus ligation 200 10,6 ± 3,2 *** 54,3 9,6 ± 0,8 OMP + pylorus ligation 20  3,9 ± 0,5 *** 83,2 3,4 ± 0,4 *** FAMC + pylorus ligation 25  5,5 ± 0,7 *** .sup.Oab 76,3 9,5 ± 0,7 FAMC + pylorus ligation 200  0,3 ± 0,3*** °ab 98,7 9,4 ± 0,9 FAMC: Suspension of fatty alcohols plus microcrystalline cellu ose MC: Microcrystalline cellulose suspension FA: Suspension of Fatty Alcohols RAT: Fatty alcohol tablet registered as a nutritional supplement I inhibition, Data as Mean ± SEM (standard error of the mean) * p < 0.05, ** p < 0.01, *** p < 0.0001, comparison vs positive control ° p < 0.0001 comparison vs similar dose of FA, a p < 0.0001 comparison vs similar doses of MC b p < 0.0001 comparison vs similar doses of RAT (Mann Whitney U test)

[0036] As a conclusion of the test presented in this example, it can be stated that oral administration with the FAMC suspension markedly and significantly inhibited the formation of gastric ulcer induced by ligation of the pylorus in rats, with a higher efficacy than that achieved with the registered tablet and with FA and MC suspensions at the same doses. Additionally, it was observed that the efficacy of the FAMC suspension was greater than the sum of the efficacy of the FA and MC suspensions in both doses evaluated, which demonstrates the existence of a synergistic effect in the composition object of the present invention.

EXAMPLE 6

[0037] The composition obtained in example 1 (FAMC) was subjected to a preclinical test with animals, in which its chemoprotective effect was compared with the chemoprotective effects of the registered fatty alcohol tablet (RAT), and suspensions of Fatty Alcohols (FA) and microcrystalline cellulose (MC) at equal concentrations to those present in the FAMC composition. For the evaluation, colon cancer was induced to Holtzmann rats with 1,2-dimethylhydrazine (DMH). The animals, with three months of age and 250±20 g; were randomly distributed into the following groups: sodium polysorbate at 2 mL/kg; DMH 20 mg/kg; and DMH plus FAMC, RAT, FA; and MC suspensions.

[0038] DMH was prepared at 4 mg/mL in distilled water, with 0.4 mg/mL EDTA as stabilizer at pH 6.5. It was administered subcutaneously at 20 mg/kg body weight once a week for 18 weeks, while the suspensions and polysorbate were administered orally for the 18-week study period. After the scheduled time, one hour after the last administration, the rats were sacrificed with 100 mg/kg pentobarbital, and the colon was removed for histopathological analysis, it was gently washed with saline solution to remove blood and adhering debris to the tissue; the tumor mass areas were fixed in a 10% buffered formalin solution for 7 days; then, parts of 3-5 μm were selected and fixed in paraffin and stained with hematoxylin and eosin. The evaluation was based on microscopic observations: neoplasia and dysplasia. The statistical analysis to evaluate the chemoprotective effect was the Shapiro-Wilk normality test, the non-parametric Kruskal Wallis and the Fisher tests with a 95% confidence level.

[0039] Table 4 summarizes the percentage of histopathological observations of the colon of the rats included in the study, induced with colon cancer and treated with the different suspensions. When applying the Kruskal Wallis test, with a confidence level of 95%, a p<0.05 was found in all cases, the results of the different groups show significant differences between all types of treatment and the phase of cancer development (normal, dysplasia and neoplasia). All animals treated only with DMH (100%) developed dysplasia or neoplasia, which shows the functioning of the model. The only group where the appearance of neoplasia was not observed was the one treated with the FAMC composition, and only 20% of these animals developed dysplasia, which implies that 80% of these animals were not affected by the application of the carcinogenic agent.

[0040] In the group treated with the suspensions FA, MC and RAT, only the 20, 10 and 10% of the animals, respectively, were not affected by the cancer inducing agent. In this way, it was demonstrated a relevant synergistic effect between the extract of alcohols and the MC, which gives a chemoprotective effect to the suspension, object of the present invention. Chemoprotective effect of FAMC suspension was much greater that the chemoprotective effect of the suspensions of such individual substances.

TABLE-US-00004 TABLE 4 Histopathological results in the colon of rats treated with vehicle (PS), dimethylhydrazine (DMH) and with different suspensions (frequency percentage). Histopathological Result Treatment Frecuency (%) No Changes (normal) Vehicle (polysorbate solution) 100 DMH   0+++ DMH + suspension FAMC  80 DMH + suspension RAT  10++ DMH + suspension FA  20++ DMH + suspension MC  10++ Dysplasia Vehiculo (polysorbate solution)   0 DMH  30+ DMH + suspension FAMC  20 DMH + suspension RAT  40+ DMH + suspension FA  50++ DMH + suspension MC  60++ Neoplasia Vehiculo (polysorbate solution)  0 DMH  70+++ DMH + suspension FAMC  0 DMH + suspension RAT  50++ DMH + suspension FA  30+ DMH + suspensoon MC  30+ PS: 3% sodium polysorbate. Statistical evaluation was performed using the Kruskal-Wallis test to determine the distribution in each cancer stage group with respect to treatment categories; significance level of 0.05. Normal (p = 0.01), dysplasia (p = 0.049), neoplasia (p=0.01). + p < 0.05; ++ p < 0.01; +++ p < .001 compared to the group with polysorbate vehicle; (Fisher test).

[0041] In conclusion, the indicators of the histopathological study revealed that the suspension containing the mixture of fatty alcohols and microcrystalline cellulose (FAMC) was the only one that demonstrated significant antitumor activity in colon cancer induced by dimethylhydrazine in rats with 100% inhibition of neoplasia.