Patchouli alcohol, patchouli alcohol compound antimalarial composition, and use in preparation of Anti-plasmodial drug

Abstract

The present invention relates to the technical field of drug application. Disclosed are patchouli alcohol, a patchouli alcohol compound antimalarial composition, and a use in preparation of an anti-plasmodial drug. The antimalarial pharmaceutical composition is a composition of artesunate and patchouli alcohol. By compounding patchouli alcohol and artesunate, synergistic antimalarial activity can be produced; and by reducing the plasmodium infection rate in red blood cells, patchouli alcohol and the compound antimalarial composition alleviate edema and inflammation in the liver and the spleen caused by malaria, and decrease haemozoin deposition in the liver and the spleen, having minimal toxic and side effects on the liver while exerting the antimalarial activity. The antimalarial effect of patchouli alcohol is obviously superior to that of patchouli oil, the antimalarial effect of the composition of artesunate and patchouli alcohol is obviously superior to that of artesunate or patchouli alcohol used alone, and patchouli alcohol and the composition of artesunate and patchouli alcohol especially have a more obvious inhibitory effect on artesunate-resistant plasmodium.

Description

Patchouli alcohol, Patchouli alcohol compound antimalarial composition and application in preparation of antimalarial parasite drugs Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to patchouli alcohol, a patchouli alcohol compound antimalarial composition and application thereof in the preparation of antimalarial protozoan drugs. Background Art

[0002] Malaria, a vector-borne infectious disease caused by Plasmodium, poses a serious threat to human health and life. Typical symptoms of malaria include fever, chills, fatigue, vomiting, and headache; severe cases can cause cerebral malaria, acute renal failure, and respiratory distress. When the density of Plasmodium in an infected individual's blood reaches a fever threshold, hemolysis of infected red blood cells, the release of merozoites and other malarial antigens, and the resulting inflammatory response trigger a malarial attack, characterized by symptoms such as high fever and chills. Sometimes, due to extensive hemolysis and liver damage caused by the deposition of hemochromatin, patients experience hepatomegaly, tenderness, and jaundice. Splenomegaly is also a common clinical manifestation of malaria, primarily due to splenic congestion and mononuclear-macrophage hyperplasia.

[0003] The discovery and application of artemisinin have enabled significant progress in malaria control and eradication. Artemisinin combination therapy (ACT) has become the most effective antimalarial treatment recommended by the World Health Organization. Artesunate is the first-line treatment in ACT and the preferred drug for treating severe malaria, including cerebral malaria. It also has some anti-inflammatory properties. However, the relapse rate is high when treated with artesunate alone. In recent years, there have been reports of varying degrees of reduced efficacy in the combination of artemisinin with quinolines, antifolates, and other drugs. This decline in efficacy is primarily due to the development of resistance to adjuvant drugs in ACT, which reduces parasite susceptibility and significantly increases relapse rates. One approach to addressing resistance is to extend the duration of treatment or to replace adjuvant drugs in ACT to which resistance has developed. If parasites develop widespread resistance to adjuvant drugs, humanity will face a potential shortage of available drugs. Therefore, the search for new antimalarial drugs and adjuvant drugs in ACT is of great clinical significance.

[0004] Previous studies have found that patchouli essential oil (patchouli oil) has antimalarial effects, but this effect is limited. Increased doses can cause significant liver and heart toxicity, leading to death in mice in severe cases, a key factor hindering its application. Patchouli essential oil has a relatively complex composition, primarily consisting of polycyclic aromatic hydrocarbons, ketones, alcohols, and terpenoids, as well as some alkaloids. The specific components or components that exert antimalarial effects are unknown, hindering its application in the antimalarial field.

[0005] Patchouli alcohol is a tricyclic sesquiterpenoid compound with the molecular formula C 15 H 26 O, molecular weight 222.37, colorless crystals. Insoluble in water, soluble in ethanol, petroleum ether and other common organic solvents.

[0006]

[0007] Baiqiu Lichun

[0008] Modern pharmacological research shows that Baiqiuli alcohol has the effects of preventing and treating liver fibrosis, kidney damage, chronic gastritis, Alzheimer's disease, anti-tumor, antibacterial, antiviral (influenza A, type B, avian influenza, Coxsackie, adenovirus), and treating gastric motility disorders and male sexual dysfunction.

[0009] At present, there is no application of Patchouli alcohol in the treatment of malaria, nor is there any research on antimalarial drug compositions related to Patchouli alcohol. Summary of the Invention

[0010] In view of the widespread drug resistance of artemisinin drugs and auxiliary drugs in the existing "artemisinin combination therapy" technology, there is an urgent need to discover new antimalarial drugs for "artemisinin combination therapy". The present invention provides patchouli alcohol, patchouli alcohol composite antimalarial composition and the use of it in the preparation of antimalarial parasite drugs. Patchouli alcohol and its composition exert their antimalarial effects by reducing the malarial parasite infection rate in red blood cells, alleviating edema and inflammation of the liver and spleen caused by malaria, and reducing heliotropin deposition in the liver and spleen.

[0011] The present invention is achieved through the following technical solutions:

[0012] A patchouli alcohol compound antimalarial composition, which is a composition of artesunate and patchouli alcohol; the mass ratio of the artesunate to the patchouli alcohol is 0.5-100:1-100.

[0013] Furthermore, the mass ratio of artesunate to patchouli alcohol is 1.73:5.

[0014] Furthermore, the patchouli alcohol compound antimalarial composition also includes at least one of a diluent, a binder, an absorbent, a disintegrant, a dispersant, a wetting agent, a solubility aid, a buffer and a surfactant.

[0015] The present invention discloses a patchouli alcohol compound antimalarial composition or application of patchouli alcohol in the preparation of antimalarial protozoan drugs.

[0016] Furthermore, the malarial parasite is Plasmodium berghei, preferably an artesunate-resistant strain of Plasmodium berghei.

[0017] Furthermore, the compound antimalarial pharmaceutical composition further comprises at least one of a diluent, a binder, an absorbent, a disintegrant, a dispersant, a wetting agent, a solubilizing agent, a buffer and a surfactant.

[0018] Patchouli oil has some effectiveness in treating malaria, but the high concentration required (2 mL / kg) limits its use in the antimalarial field, resulting in significant toxicity. Research has shown that the patchouli alcohol component in patchouli oil can reduce the infection rate of Plasmodium in red blood cells, alleviate malaria-induced edema and inflammation in the liver and spleen, and reduce heliotropin in the liver and spleen, exerting antimalarial effects. The drug has a particularly pronounced inhibitory effect against artesunate-resistant Plasmodium.

[0019] Research has found that at a dose of 5-100 mg / kg, Patchouli alcohol can produce an inhibitory effect on Plasmodium berghei and artesunate-resistant strains of Plasmodium berghei. However, for patchouli oil (patchouli alcohol content 27.03%), a dose of 2 mL / kg can only achieve a weak antimalarial effect. This further shows that patchouli oil not only contains components that produce liver toxicity, but also likely contains components that have antagonistic effects with Patchouli alcohol, affecting the antimalarial effect of Patchouli alcohol.

[0020] Research has also found that the combined administration of patula alcohol and artesunate can produce synergistic antimalarial activity, inhibit Plasmodium berghei (especially artesunate-resistant strains of Plasmodium berghei), reduce the infection rate of Plasmodium in red blood cells, alleviate edema and inflammation of the liver and spleen caused by malaria, and reduce malarial pigment deposition in the liver and spleen to exert an antimalarial effect.

[0021] The beneficial effects achieved by the present invention are:

[0022] Patchouli alcohol and the Patchouli alcohol compound antimalarial composition reduce the infection rate of Plasmodium in red blood cells, alleviate the edema and inflammation of the liver and spleen caused by malaria, and reduce malarial pigment deposition in the liver and spleen, thereby exerting antimalarial activity with minimal toxic side effects on the liver; Patchouli alcohol and the Patchouli alcohol compound antimalarial composition have a more obvious inhibitory effect on artesunate-resistant Plasmodium, especially the composition can produce synergistic antimalarial activity, solve the problem of Plasmodium resistance, have good application effect, and have important clinical significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] FIG1 is a graph showing changes in body weight of malarial mice in different groups during drug administration in Example 2;

[0024] Figure 2 shows the Giemsa staining results (1000×) of different groups of malaria-infected mice in Example 2 24 hours after the last administration, including group B (a), group M (b), group SA (c), group PA (d), and group PA+SA (e);

[0025] FIG3 is a graph showing the changes in infection rates of different groups of malaria-stricken mice during drug administration in Example 2;

[0026] FIG4 is a graph showing the infection rates of different groups of malaria-stricken mice 24 hours after the last administration of the drug in Example 2;

[0027] Figure 5 shows images of the liver and spleen of different groups of malaria-infected mice 24 hours after the last administration of the drug in Example 2, including group B (a), group M (b), group SA (c), group PA (d), and group PA+SA (e);

[0028] FIG6 is a graph showing liver indexes of different groups of malarial mice 24 hours after the last administration in Example 2;

[0029] FIG7 is a graph showing spleen index of different groups of malaria-stricken mice 24 hours after the last administration in Example 2;

[0030] Figure 8 shows HE-stained liver tissue sections of different groups of malaria-stricken mice 24 hours after the last administration of the drug in Example 2 (magnifications are 40×, 100×, and 400×, respectively), including group B (a), group M (b), group SA (c), group PA (d), and group PA+SA (e);

[0031] Figure 9 shows HE-stained spleen tissue sections of different groups of malaria-stricken mice 24 hours after the last administration of the drug in Example 2 (magnifications are 40×, 100×, and 400×, respectively), including group B (a), group M (b), group SA (c), group PA (d), and group PA+SA (e). DETAILED DESCRIPTION

[0032] The following further describes the above content of the present invention in detail through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention.

[0033] Patchouli alcohol in the present invention was purchased from the China Food and Drug Administration, batch number: 111822-201904; artesunate was purchased from Yuanye Biotechnology (Shanghai, China), batch number: X03511Y123220. Physiological saline, 4% paraformaldehyde, Tween 80, phosphate buffer solution, and methanol were used.

[0034] Patchouli is a medicinal herb grown in Hainan. It is the whole herb of Pogostemon cablin (Blanco) Benth, a plant of the genus Pogostemon in the family Lamiaceae. Ten kilograms of the dried aerial parts of the herb were extracted using a steam distillation apparatus. The volatile oil was then dried over anhydrous sodium sulfate to remove water, yielding 53 g of patchouli volatile oil (patchouli oil). The resulting patchouli oil was analyzed using an HP-5 capillary column (30 m length, 0.25 mm inner diameter) according to the method specified in the 2020 edition of the Chinese Pharmacopoeia. The resulting patchouli oil was found to contain 27.03% patchouli alcohol, which meets the requirements of the Chinese Pharmacopoeia.

[0035] SPF female KM mice (18 ± 2) g were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. (license number: SCXK (Beijing) 2016-0011). All mice were housed in the animal room of Shandong Institute of Traditional Chinese Medicine with free access to food and water. The mice were housed under a 12-h light and 12-h dark cycle at room temperature of (22 ± 2)°C and a relative humidity of 55 ± 5%. Water, bedding, and feed were changed regularly. Bedding and feed were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd. (production license number: SCXK (Lu) 20190003). The experimental procedures were strictly in accordance with the animal care regulations.

[0036] Giemsa staining: After inoculation with Plasmodium, peripheral blood was collected from mice for smears. All smears were fixed with methanol and stained with Giemsa for 20 minutes. The level of malarial parasitemia in each mouse's peripheral blood was examined microscopically. Finally, the following formulas were used: infection rate (%) = [(total number of infected red blood cells counted / total number of red blood cells counted) × 100%), and inhibition rate (%) = (average infection rate in the model group - malarial parasite infection rates in mice in the other treatment groups) / average infection rate in the model group × 100%.

[0037] Hematoxylin and eosin staining: 24 h after the last administration, the mice were anesthetized, the abdominal cavity was opened, and the liver and spleen of the mice were separated and weighed. After being fixed in 4% paraformaldehyde at room temperature for ≥ 24 h, they were embedded in paraffin and sectioned. The sections were stained with hematoxylin and eosin to determine the changes in the structure of the liver and spleen.

[0038] Statistical methods: GraphPad Prism 8.0.2 statistical software was used for statistical tests. Measurement data were expressed as means. One-way analysis of variance (ANOVA) was used to compare means among multiple groups. Pairwise comparisons were performed using the LSD method after the groups met the requirements of normality and homogeneity of variance. P < 0.05 indicated statistically significant differences.

[0039] Example 1

[0040] Inhibitory effects of different doses of Patchouli alcohol on Plasmodium berghei

[0041] (1) Animal grouping and drug administration

[0042] Animal model establishment: SPF female KM mice (18 ± 2 g) were fed adaptively for 3 days and each mouse was inoculated with 1 × 10 7 A red blood cell infected with Plasmodium berghei.

[0043] Mice infected with Plasmodium berghei were randomly divided into a model group (group M, ig 3% Tween 80, ip normal saline), a low-dose Patchouli alcohol group (low-dose PA group, ig 5 mg / kg PA, ip normal saline), a medium-dose Patchouli alcohol group (medium-dose PA group, ig 20 mg / kg PA, ip normal saline), a high-dose Patchouli alcohol group (high-dose PA group, ig 100 mg / kg PA, ip normal saline), and a positive control artesunate group (group SA, ig 3% Tween 80, ip 1.73 mg / kg SA). A blank group (group B, ig 3% Tween 80, ip normal saline) was also set up. The ig and ip administration volume was 0.2 mL / 20 g. There were 6 mice in each group. Dosing began 2 hours after inoculation on the day of inoculation and continued for 4 consecutive days. The mental state of the mice was observed and the body weight was recorded before dosing every day.

[0044] (2) Infection rate and inhibition rate detection

[0045] 24 hours after the last administration, the infection rate and inhibition rate were calculated using the Giemsa staining method. As shown in Table 1 below, 5-100 mg / kg doses of patchouli alcohol all had inhibitory effects on Plasmodium berghei.

[0046] Table 1 Infection rate and inhibition rate of Plasmodium infection in different groups of mice 24 hours after the last administration (n=6)

[0047]

[0048] Note: * P < 0.05, compared with group M; # P<0.05, compared with SA group

[0049] Example 2

[0050] The inhibitory effect of Patchouli alcohol and Patchouli alcohol combined antimalarial composition on Plasmodium berghei.

[0051] (1) Animal grouping and drug administration

[0052] Animal model establishment: SPF female KM mice (18 ± 2 g) were fed adaptively for 1 week and each mouse was inoculated with 1×10 7 The mice were inoculated with red blood cells infected with Plasmodium berghei. On the second day of inoculation, the tails were cut and blood smears were taken to observe the infection status of the mice.

[0053] Mice infected with Plasmodium berghei were randomly divided into a model group (M group, 3% Tween 80 administered intravenously and saline solution intravenously), a patchouli alcohol group (PA group, 30 mg / kg PA administered intravenously and saline solution intravenously), an artesunate group (SA group, 3% Tween 80 administered intravenously and saline solution intravenously), and a patchouli alcohol + artesunate group (PA+SA group, 30 mg / kg PA administered intravenously and saline solution intravenously). A blank control group (Group B, 3% Tween 80 administered intravenously and saline solution intravenously) was also established. Drug administration began on the second day of inoculation and continued for four consecutive days, with a volume of 0.2 mL / 20 g administered intravenously and intravenously. Before daily administration, the mice were observed for mental status, body weights were recorded, and tail clipping and blood samples were collected to assess infection status. Mice in Group B were agile, with natural, lustrous fur. There were no significant abnormalities in diet, drinking water, or feces. Compared with the other groups, weight gain was faster. The other infected mice showed varying degrees of decreased exploratory behavior and a decreased response to external stimuli by the third day of administration. The changes in body weight of mice in different groups during the drug administration period are shown in Figure 1. Compared with group B, the body weight of mice in group M decreased significantly from the third day, while the body weight of mice in groups SA and PA decreased slowly.

[0054] (2) Infection rate and inhibition rate detection

[0055] 24 hours after the last administration, the infection rate and inhibition rate were calculated by Giemsa staining. The results are shown in Table 2 below. The Giemsa staining results of mice in different groups are shown in Figure 2. The changes in infection rates of malaria-infected mice in different groups during administration are shown in Figure 3. The infection rates of malaria-infected mice in different groups 24 hours after the last administration are shown in Figure 4.

[0056] As shown in Figure 2, the M group contained a large number of late trophozoites and ring masses, and the red blood cell structures were more broken. The above situation was significantly alleviated in the SA group and the PA+SA group. As shown in Figure 3, through continuous observation of the infection rate of mice during the drug administration period, the growth trend of the infection rate of mice in the SA group and the PA+SA group was significantly slowed down (P<0.05). Compared with the SA group, the infection rate of the PA+SA group was significantly reduced (P<0.05). As shown in Figure 4, 24 hours after the last administration, there was a great difference in the infection rate among mice in each group.

[0057] Table 2 Infection rate and inhibition rate of Plasmodium infection in different groups of mice 24 hours after the last administration (n=6)

[0058]

[0059] Note: * P < 0.05, compared with group M; # P<0.05, compared with SA group

[0060] (3) Histopathological analysis

[0061] Observation of the liver and spleen of the different groups of malaria-infected mice 24 hours after the final administration (as shown in Figure 5) revealed that compared with Group B, all mice exhibited enlarged and darker livers and spleens. The swollen organs were fragile and easily fractured during experimental manipulation. This condition was alleviated in the SA and PA+SA groups, with the alleviation being more pronounced in the PA+SA group. As shown in Figures 6 and 7, there were no significant differences in liver and spleen indices among the SA, PA, and PA+SA groups.

[0062] HE-stained liver tissue sections of the different groups of malaria-infected mice 24 hours after the last administration are shown in Figure 8. The hepatocyte morphology of the mice in group B was normal, the cell nucleus was centrally located, and the hepatocytes were arranged regularly and densely. In contrast, obvious inflammation, cell necrosis, and damage were observed in the liver of the mice in group M. The hepatocytes were uneven in size, swollen, and loose in tissue. Vacuole-like lesions appeared in the hepatocytes, and a large amount of hemochromatin was deposited. All the above symptoms were significantly alleviated in the SA group, PA group, and PA+SA group.

[0063] HE-stained spleen tissue sections of the different groups of malaria-infected mice 24 hours after the last administration are shown in Figure 9. The spleen tissue in group M was severely congested and the red pulp was enlarged. The white pulp structure was destroyed, the clear edge area around the follicles became unclear, the red pulp was widely vacuolated, and a large amount of hemochromatin was deposited. All the above symptoms in the SA, PA, and PA+SA groups were relieved, and the improvement of the above symptoms in the PA+SA group was more significant.

[0064] Example 3

[0065] The inhibitory effect of different doses of Patchouli alcohol combined with antimalarial composition on Plasmodium berghei.

[0066] Animal model establishment: SPF female KM mice (18 ± 2 g) were fed adaptively for 3 days and each mouse was inoculated with 1 × 10 7 The mice were inoculated with red blood cells infected with Plasmodium berghei. On the second day of inoculation, the tails were cut and blood smears were taken to observe the infection status of the mice.

[0067] Mice infected with Plasmodium berghei were randomly divided into a model group (M group, ig 3% Tween 80, ip normal saline), a patchouli alcohol group (PA group, ig 100 mg / kg PA, ip normal saline), an artesunate group (SA group, ig 3% Tween 80, ip 1.73 mg / kg SA), a low-dose patchouli alcohol + artesunate combination group (ig 5 mg / kg PA, ip 1.73 mg / kg SA), a medium-dose patchouli alcohol + artesunate combination group (ig 20 mg / kg PA, ip 1.73 mg / kg SA), and a high-dose patchouli alcohol + artesunate combination group (ig 100 mg / kg PA, ip 1.73 mg / kg SA), with 6 mice in each group. Drugs were administered starting on the second day of inoculation and for 4 consecutive days, with an ig and ip administration volume of 0.2 mL / 20 g. The mental state of the mice was observed and their body weight was recorded before each day of drug administration. Blood was collected by tail clipping to check the infection status of the mice.

[0068] Twenty-four hours after the last administration, infection rates and inhibition rates were calculated using Giemsa staining. The results are shown in Table 3. Compared with the M group, PA alone had a modest inhibitory effect on P. berghei. SA alone, and different doses of PA (5 mg / kg, 20 mg / kg, and 100 mg / kg) and SA combined had significant inhibitory effects on P. berghei compared with the M group (P < 0.05). The infection rate of mice treated with the PA + SA combination was significantly lower than that of mice treated with SA alone, indicating that the PA + SA combination was more effective than SA alone. The inhibition rate in the PA + SA combination group was approximately 12% higher than that in the SA alone group and greater than the combined inhibition rates of SA and PA alone (approximately 50%), indicating that PA has a synergistic effect on SA in killing P. berghei. There was no significant difference in infection rate between the different doses of PA (low, medium, and high) administered in combination with SA. Therefore, the optimal dose of PA for combined use with SA is 5 mg / kg.

[0069] Table 3 Infection rate and inhibition rate of Plasmodium infection in different groups of mice 24 hours after the last administration (n=6)

[0070]

[0071] Note: * P < 0.05, compared with group M; # P<0.05, compared with SA group

[0072] Example 4

[0073] Inhibitory effect of Patchouli alcohol on artesunate-resistant Plasmodium

[0074] (1) Animal grouping and drug administration

[0075] Animal model establishment: SPF female KM mice (18 ± 2 g) were fed adaptively for 3 days and each mouse was inoculated with 1 × 10 7 Red blood cells infected with artesunate-resistant Plasmodium falciparum.

[0076] Mice infected with artesunate-resistant Plasmodium were randomly divided into a model group (RM group, ig administration of 3% Tween 80, ip administration of normal saline), a patchouli alcohol group (R-PA group, ig administration of 5 mg / kg patchouli alcohol, ip administration of normal saline), a patchouli oil group (R-PO group, ig administration of 2 mL / kg PO, ip administration of normal saline), a positive control artesunate group (R-SA group, ig administration of 3% Tween 80, ip administration of 1.73 mg / kg artesunate), and a blank control group (B group, ig administration of 3% Tween 80, ip administration of normal saline), with six mice in each group. Drug administration began on the second day after inoculation and continued for four consecutive days, with a volume of 0.2 mL / 20 g administered ig or ip. The mental state of the mice was observed and their body weight was recorded daily before drug administration. Mice in group B were agile, with natural, lustrous fur. There were no significant abnormalities in diet, drinking water, or feces. Compared with the other groups, mice in group B showed faster weight gain. The other infected mice showed varying degrees of decreased exploratory behavior and a decreased response to external stimuli by the third day of drug administration.

[0077] 24 h after the last administration, the infection rate and inhibition rate were calculated using the Giemsa staining method. The results are shown in Table 4.

[0078] Table 4 Infection rate and inhibition rate of Plasmodium infection in different groups of mice 24 hours after the last administration (n=6)

[0079]

[0080] Note: * P < 0.05, compared with group M; # P<0.05, compared with SA group

[0081] Example 5

[0082] Inhibitory effect of a patchouli alcohol-compound antimalarial composition on artesunate-resistant Plasmodium.

[0083] Animal model establishment: SPF female KM mice (18 ± 2 g) were fed adaptively for 3 days and each mouse was inoculated with 1 × 10 7 Red blood cells infected with artesunate-resistant Plasmodium falciparum.

[0084] Mice infected with artesunate-resistant Plasmodium were randomly divided into a model group (RM group, ig 3% Tween 80, ip normal saline), a patchouli alcohol group (R-PA group, ig 5 mg / kg PA, ip normal saline), an artesunate group (R-SA group, ig 3% Tween 80, ip 1.73 mg / kg SA), and a patchouli alcohol + artesunate combination group (R-PA+SA, ig 5 mg / kg PA, ip 1.73 mg / kg SA), with 6 mice in each group. The drugs were administered on the second day after inoculation for 4 consecutive days, with an ig and ip administration volume of 0.2 mL / 20 g. The mental state of the mice was observed before each administration, and their body weight was recorded. The infection status of the mice was checked by tail clipping and blood was collected.

[0085] Twenty-four hours after the last administration, the infection rate and inhibition rate were calculated using Giemsa staining. The results are shown in Table 5. Compared with the RM group, the infection rate of malaria-infected mice receiving PA alone was slightly reduced. Compared with the RM group, the infection rate of mice in the R-SA group and the R-PA + SA combination group was significantly reduced. The infection rate in the R-PA + SA group was also slightly lower than that in the R-SA group. The inhibition rate in the R-PA + SA combination group (53.76%) was nearly 20% higher than that in the R-SA alone group (34.54%) and greater than the combined inhibition rates of R-SA and R-PA alone (approximately 48%), indicating that PA has a synergistic effect on SA in killing artesunate-resistant Plasmodium parasites.

[0086] Table 5 Infection rate and inhibition rate of Plasmodium infection in different groups of mice 24 hours after the last administration (n=6)

[0087]

[0088] Note: * P < 0.05, compared with the RM group; # P<0.05, compared with R-SA group

[0089] Example 6

[0090] Inhibitory effects of different doses of patchouli oil on Plasmodium berghei.

[0091] Animal model establishment: SPF female KM mice (18 ± 2 g) were fed adaptively for 1 week and each mouse was inoculated with 1×10 7 The mice were inoculated with red blood cells infected with Plasmodium berghei. On the second day of inoculation, the tails were cut and blood smears were taken to observe the infection status of the mice.

[0092] Mice infected with Plasmodium berghei were randomly divided into a model group (3% Tween 80 administered intravenously and saline solution intravenously), a low-dose patchouli oil group (1 mL / kg intravenously and saline solution intravenously), a medium-dose patchouli oil group (2 mL / kg intravenously and saline solution intravenously), and a high-dose patchouli oil group (3 mL / kg intravenously and saline solution intravenously). A blank control group (3% Tween 80 administered intravenously and saline solution intravenously) was also included. Six mice were administered the drug starting on the second day of inoculation and continued for four consecutive days, with a volume of 0.2 mL / 20 g administered intravenously and intravenously. Before each daily dose, the mice were observed for mental status, body weights were recorded, and tails were cut to bleed for infection status. The study found that mice died after receiving a 3 mL / kg dose of patchouli oil. Patchouli oil at a 2 mL / kg dose exhibited some antimalarial activity, while a 1 mL / kg dose had no antimalarial effect.

Claims

1. A Patchouli alcohol composite antimalarial composition, characterized in that: The patchouli alcohol composite antimalarial composition is a composition of artesunate and patchouli alcohol, and the mass ratio of artesunate to patchouli alcohol is 0.5-100: 1-100.

2. The Patchouli alcohol composite antimalarial composition according to claim 1, characterized in that: The mass ratio of artesunate to patchouli alcohol is 1.73:

5.

3. The Patchouli alcohol composite antimalarial composition according to claim 1, characterized in that: The patchouli alcohol composite antimalarial composition also includes at least one of a diluent, a binder, an absorbent, a disintegrant, a dispersant, a wetting agent, a cosolvent, a buffer and a surfactant.

4. Use of the Patchouli alcohol composite antimalarial composition according to any one of claims 1 to 3 in the preparation of an antimalarial protozoan drug, characterized in that: The malarial parasite is Plasmodium berghei.

5. A use of Patchouli alcohol in the preparation of an anti-malarial drug, characterized in that: The malarial parasite is Plasmodium berghei.

6. Use of the Patchouli alcohol composite antimalarial composition according to claim 4 in the preparation of antimalarial parasite drugs, characterized in that: The Plasmodium berghei is an artesunate-resistant strain of Plasmodium berghei.

7. Use of the Patchouli alcohol composite antimalarial composition according to claim 4 in the preparation of an antimalarial parasite drug, characterized in that: The composite antimalarial drug composition also includes at least one of a diluent, a binder, an absorbent, a disintegrant, a dispersant, a wetting agent, a solubilizing agent, a buffer and a surfactant.

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