Medicinal composition of plant aromatic essential oil and application of medicinal composition

Abstract

The invention discloses a pharmaceutical composition of plant aromatic essential oil and application of the pharmaceutical composition. The invention provides a pharmaceutical composition. The pharmaceutical composition comprises beta-caryophyllene, alpha-humulene and caryophyllene oxide. The pharmaceutical composition can obviously improve the pain threshold value and reduce the level of inflammatory factors in cells, and can be used for treating neuropathy caused by chemotherapeutic drugs.

Description

Technical Field

[0001] The invention relates to a plant aromatic essential oil medicinal composition and application thereof. Background Art

[0002] With the progress of cancer diagnosis and treatment, the survival time of many patients will continue to improve, and peripheral neuropathy caused by cancer and chemotherapy has attracted more and more attention from patients and their families. Among them, peripheral neuropathy caused by chemotherapy is an adverse reaction with a high incidence and long duration.

[0003] Paclitaxel, as a representative plant molecule used in first-line chemotherapy, still attracts wide attention. However, chemotherapy drugs represented by this type of drug often have adverse reactions such as bone marrow suppression, nervous system damage, digestive system toxicity, and blood system damage. The mechanisms of these reactions include microtubule destruction, oxidative stress, mitochondrial damage, changes in ion channel activity, immune processes, and neuroinflammation. The main symptoms are symmetrical numbness, abnormal touch, and pain in the distal extremities, accompanied by loss of tendon reflexes and movement disorders. As a part of cancer pain, neuropathic pain caused by chemotherapy seriously affects the survival and prognosis of patients.

[0004] Cannabis is an annual herbaceous plant of the genus Cannabis in the family Moraceae, Urticales, class dicotyledons. Hemp with a content of less than 0.3% of the psychoactive ingredient tetrahydrocannabinol is called industrial hemp, of which cannabinoids and volatile oils are the main active ingredients with rich pharmacological activity. Currently, there have been many studies on cannabinoids and there are already drugs on the market, but there is less use of volatile oil components. Summary of the invention

[0005] The technical problem solved by the present invention is to provide a plant essential oil pharmaceutical composition and its application. The plant essential oil of the present invention contains active substances β-caryophyllene, α-humulene and caryophyllene oxide. Furthermore, the plant essential oil of the present invention has one or more of the following advantages: the plant essential oil has a synergistic effect, can be used to treat peripheral neuropathy caused by tumor chemotherapy drugs, and has good application prospects.

[0006] The present invention solves the above technical problems through the following technical solutions.

[0007] The present invention provides a pharmaceutical composition, which comprises: beta-caryophyllene, alpha-humulene and caryophyllene oxide.

[0008] In some embodiments, the pharmaceutical composition further comprises one or more of (-)-humulene epoxide II, α-bisabolol and (+)-delta-cadinene, cnidium-3,7(11)-diene, γ-ylang-oleene, hordein, β-thulene, α-bergamotene, and bergamotene oxide.

[0009] In some embodiments, in the pharmaceutical composition, the content of β-caryophyllene is preferably 10-35%; more preferably 15-30%; for example 26.49%.

[0010] In the pharmaceutical composition, the content of α-humulene is preferably 5-20%, more preferably 10-15%, for example 12.11%.

[0011] In the pharmaceutical composition, the content of caryophyllene oxide is preferably 8-20%, more preferably 10-15%, for example 11.65%.

[0012] In the pharmaceutical composition, the content of the (-)-humulene epoxide II is preferably 3-5%, for example 3.76%.

[0013] In the pharmaceutical composition, the content of α-bisabolol is preferably 2-5%, for example 3.27%.

[0014] In the pharmaceutical composition, the content of (+)-δ-cadinene is preferably 2-5%, for example 3.24%.

[0015] In the pharmaceutical composition, the content of cnidium monnieri-3,7(11)-diene is preferably 2-5%, for example 3.07%.

[0016] In the pharmaceutical composition, the content of γ-ylang-ylangolene is preferably 2-4%, for example 2.91%.

[0017] The hordein content is preferably 2-4%, for example 2.83%.

[0018] The β-surene content is preferably 2-4%, for example 2.44%.

[0019] The α-bergamotene content is preferably 2-4%, for example 2.37%.

[0020] The content of bergamot oxide is preferably 1-3%, for example 1.88%.

[0021] In some embodiments, the pharmaceutical composition satisfies one or more of the following:

[0022] (1) The active ingredients of the pharmaceutical composition are derived from plant essential oils;

[0023] (2) The plant is industrial hemp flowers and leaves;

[0024] (3) The active ingredient of the pharmaceutical composition is mainly sesquiterpenoid compounds.

[0025] In some embodiments, the active ingredients of the pharmaceutical composition consist of active substances β-caryophyllene, α-humulene and caryophyllene oxide.

[0026] In some embodiments, the active ingredients of the pharmaceutical composition include β-caryophyllene, α-humulene and caryophyllene oxide, (-)-humulene epoxide II, α-bisabolol and (+)-delta-cadinene, cnidium-3,7(11)-diene, γ-ylang-ylangolene, hordein, β-thurene, α-bergamotene and bergamotene oxide.

[0027] In some embodiments, the pharmaceutical composition is prepared by the following method, which comprises the following steps: (1) extracting and collecting volatile oil from industrial hemp flower and leaf powder by steam distillation;

[0028] (2) subjecting the collected volatile oil to gas phase separation to obtain the pharmaceutical composition.

[0029] Preferably, the solvent of the steam distillation method is water.

[0030] The distillation temperature of the steam distillation method is 150°C to 170°C; for example, 160°C.

[0031] The gas phase chromatographic column is a HP-5MSUI column.

[0032] The injection port temperature of the gas phase is 200°C to 280°C, for example 250°C.

[0033] The gas phase chromatographic column flow rate is 0.8 ml / min to 12 ml / min, for example 1 ml / min.

[0034] The gas phase chromatographic column split ratio is 1:1 to 3:1, for example 2:1.

[0035] More preferably, the gas phase is programmed to heat, and the method of programmed heating is preferably: initial temperature 60°C, maintained for 2 minutes; increased to 110°C at 20°C / min; then increased to 160°C at 2°C / min, maintained for 1 minute; and then increased to 280°C at 20°C / min, maintained for 3 minutes.

[0036] The present invention also provides a use of the above-mentioned drug combination in preparing a drug for preventing and / or treating peripheral neuropathy pain.

[0037] In some embodiments, the medicament is used to increase SOD activity.

[0038] In some embodiments, the medicament is used to alleviate oxidative stress.

[0039] In some embodiments, the medicament is used to reduce sensitivity to thermal stimulation and / or mechanical stimulation.

[0040] In some embodiments, the medicament is for preventing and / or treating pain from inflammation.

[0041] In some embodiments, the peripheral neuropathy pain is mediated by inflammatory cytokines.

[0042] In some embodiments, the inflammatory cytokine may be one or more of IL-6, IL-10, TNF-α, and MCP-1.

[0043] In some embodiments, the peripheral neuropathy pain is chemotherapeutic drug-induced pain.

[0044] The present invention also provides a use of the above-mentioned drug combination in the preparation of an inhibitor, wherein the inhibitor is used to inhibit the secretion of inflammatory cytokines.

[0045] In some embodiments, the inflammatory cytokine may be one or more of IL-6, IL-10, TNF-α, and MCP-1.

[0046] As used herein, the term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) ameliorating one or more biological manifestations of the disease or condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition.

[0047] The term "pharmaceutical composition" as used herein refers to a composition containing specified active ingredients that can be prepared into the same dosage form.

[0048] The term "subject" as used herein refers to any animal that is about to or has been administered the compound or composition according to the embodiments of the present invention, preferably a mammal, and most preferably a human. As used herein, the term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being the most preferred.

[0049] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0050] Unless otherwise specified, the reagents and raw materials used in the present invention are commercially available.

[0051] The positive and progressive effects of the present invention are: the present invention provides a new plant aromatic oil drug combination obtained from industrial hemp and having multiple active substances. Furthermore, the drug combination of the present invention has one or more of the following advantages: (1) the drug combination has a synergistic effect; (2) it can be used to treat peripheral neuropathy caused by anti-tumor drugs; (3) it has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 This is the total ion current obtained by GC-MS analysis of industrial hemp volatile oil

[0053] Figure 2 The trend of mechanical sensitivity in mice

[0054] Figure 3 The change trend of thermal sensitivity in mice

[0055] Figure 4 Results of the mouse serum inflammatory factor IL-6 test

[0056] Figure 5 Results of the mouse serum inflammatory factor IL-10 test

[0057] Figure 6 Results of the mouse serum inflammatory factor TNF-α test

[0058] Figure 7 Results of the mouse serum inflammatory factor MCP-1 test DETAILED DESCRIPTION

[0059] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0060] Example 1 Preparation method of industrial hemp volatile oil

[0061] Weigh industrial hemp flower and leaf powder (raw materials provided by Yunnan Hangu Biotechnology Co., Ltd.), place it in a round-bottom flask, add pure water at a solid-liquid ratio of 8 mg / L, set the electric heating mantle to heat at 160°C and extract by steam distillation for 5 hours, collect the volatile oil, and store it in a sealed, light-proof refrigerator at 4°C.

[0062] Example 2 GC-MS analysis of industrial hemp volatile oil

[0063] Chromatography-mass spectrometry conditions: HP-5MSUI column (0.250mm×30m×0.25μm); carrier gas is high-purity helium; direct injection; injection port temperature is 250℃; transfer line temperature is 260℃; column flow rate is 1ml / min; split ratio is 2:1, ionization mode is EI, scanning range (m / z) is 50-600, and acquisition mode is full scan mode. The program temperature rise method is: initial temperature is 60℃, maintained for 2min; increased to 110℃ at 20℃ / min; then increased to 160℃ at 2℃ / min, maintained for 1min; then increased to 280℃ at 20℃ / min, maintained for 3min.

[0064] Total ion current ( Figure 1 ) were compared with reference substances and searched in the NIST database to identify the chemical components. The main components are shown in Table 1, accounting for 76% of the total peak area.

[0065] Table 1 Main chemical components of industrial hemp volatile oil

[0066]

[0067] Example 3 Study on the effect of industrial hemp volatile oil in preventing and / or treating peripheral neuropathic pain induced by chemotherapy drugs in mice

[0068] (1) Determination of basic pain threshold and grouping: Before model construction, the mice were first subjected to behavioral tests to determine their basic pain threshold. Mice with basic mechanical pain thresholds outside of 0.4-2.0g and basic thermal pain thresholds outside of 5-15s were discarded, and 70 experimental mice were included. Ten mice were randomly selected as blank controls, and the remaining 60 mice were used to construct a mouse PIPN model with paclitaxel and were divided into groups for drug administration;

[0069] (2) Modeling: The 60 mice mentioned above were given paclitaxel solution at a dose of 2 mg / kg body weight by intraperitoneal injection 1, 3, 5, and 7 days after the baseline pain threshold assessment, with a cumulative dose of 8 mg / kg body weight.

[0070] (3) Grouping and administration: The 60 model mice mentioned above were randomly divided into 6 groups, with 10 mice in each group, namely the model group, the positive drug gabapentin group (100 mg / kg, ig), the experimental drug (HEO) low, medium and high dose groups (50 mg / kg, 100 mg / kg, 150 mg / kg, ip), and the experimental drug (HEO) and AM630 combination group (HEO 100 mg / kg, ip; AM630 3 mg / kg, ip). The mice were given drug treatment every day on days 2, 4, 6, 8, 10, 12, and 14 (AM630 was administered 30 minutes before HEO administration). The blank group and the model group were intraperitoneally injected with 0.2 ml of 3% Tween80-containing saline.

[0071] (4) Observation of general physical signs: During the entire experimental process, the physical signs of the mice were observed and recorded: the weight of the mice was monitored regularly; the mice's food intake and drinking habits were observed; the mice's mental state, behavior, coat color, etc. were observed.

[0072] (5) Pain behavioral assessment - mechanical sensitivity: The mechanical sensitivity of mice was assessed one day before the first administration and on days 2, 4, 6, 8, and 14. The up-down method was used to allow the mice to adapt to the environment of the behavioral experiment and place them in a glass cover on the test bench for about 20 minutes. After the mice were quiet, 0.4-2.0g Von Frey fibers were used to mechanically stimulate the skin on the bottom of the right hind foot of the mice. Each stimulation lasted about 6 seconds, starting from 0.4g, and measured 5 times. When the number of paw withdrawal reactions was greater than 3 times, it was recorded as the last fiber stimulation intensity.

[0073] (6) Pain behavior assessment - thermal sensitivity: The mechanical sensitivity of mice was assessed one day before the first administration and on the 2nd, 4th, 6th, 8th and 14th days. The thermal analgesia was tested by placing the mice in a transparent plexiglass box. The mice were allowed to adapt to the test environment for about 20 minutes before the test. After the mice were quiet, the thermal radiation light source was moved to focus on the skin of the soles of the hind feet of the mice, and the mice's response to thermal noxious stimulation was observed. The intensity of thermal stimulation should be such that normal mice will withdraw their feet after about 5-15 seconds of exposure. To prevent tissue damage caused by prolonged thermal stimulation, the maximum duration of thermal stimulation was set to 25 seconds. The soles of the right hind feet of the mice were measured 3 times at intervals of 5 minutes. The average of the 3 withdrawal latencies was the thermal sensitivity pain threshold.

[0074] (7) Detection of inflammatory factors: Orbital blood was collected from PIPN mice on day 9 (4 mice) and day 15 (6 mice), and the supernatant was obtained by centrifugation and stored at -20°C for later use. ELISA kits were used to measure the levels of IL-6, IL-10, TNF-α, and MCP-1 in the serum.

[0075] (8) All data results are expressed as mean ± standard deviation The experimental results of each group were analyzed by T-test and one-way ANOVA using GraphPad Prism7 and SPSS software. P < 0.05 indicated statistically significant differences.

[0076] (9) Mechanical stimulation sensitivity measurement results

[0077] Starting from the second day, the mechanical sensitivity pain threshold of the model group mice decreased, which was statistically different from the blank group (P < 0.05), indicating that the modeling was effective; compared with the model group, the mechanical sensitivity pain threshold of the positive drug Gabapentin group, HEO-M group, and HEO-H group increased (P < 0.01); compared with the HEO-M group, the mechanical sensitivity pain threshold of the HEO-M combined with CB2 receptor antagonist AM630 group did not increase (P < 0.01). From the fourth day on, the mechanical sensitivity pain threshold of each drug treatment group increased, which was statistically different from the model group (P < 0.01). This shows that HEO administration can relieve the mechanical sensitivity pain of PIPN mice, which may be related to the stimulation of CB2 receptors; it is worth noting that the mechanical sensitivity pain threshold of the blank control group mice was also reduced. The reason for the analysis was that the blank group mice were sensitized due to multiple stimulations. The specific results are shown in Tables 2, 3 and Figure 2 shown.

[0078] Table 2 Mechanical sensitivity test results (expressed as Von Frey fiber final stimulation weight / g) n=10)

[0079]

[0080]

[0081] Note: Compared with the blank control group: #-P<0.05, ##-P<0.01; Compared with the model group: *-P<0.05; **-P<0.01; Compared with the HEO-M group: @@ -P<0.01.

[0082] Table 3 Mechanical sensitivity test results (expressed in Von Frey fiber final stimulation weight / g) n=10)

[0083]

[0084] Note: Compared with the blank control group: #-P<0.05, ##-P<0.01; Compared with the model group: *-P<0.05; **-P<0.01; Compared with the HEO-M group: @@ -P<0.01.

[0085] (10) Results of thermal sensitivity measurement

[0086] The thermal sensitivity pain threshold of the model group mice was measured to be reduced from the second day, which was statistically different from the blank group (P < 0.01), indicating that the modeling was effective; from the fourth day on, the thermal sensitivity pain threshold of each drug treatment group rebounded, which was statistically different from the model group (P < 0.05), indicating that HEO administration can increase the thermal sensitivity pain threshold of PIPN mice. The specific results are shown in Tables 4 and 5. Figure 3 shown.

[0087] Table 4 Results of thermal sensitivity test (expressed in mouse paw withdrawal time / s) n=10)

[0088]

[0089] Note: Compared with the blank control group: #-P<0.05, ##-P<0.01; compared with the model group: *-P<0.05; **-P<0.01.

[0090] Table 5 Results of thermal sensitivity experiment (expressed in mouse paw withdrawal time / s) n=10)

[0091]

[0092] Note: Compared with the blank control group: #-P<0.05, ##-P<0.01; compared with the model group: *-P<0.05; **-P<0.01. (11) Results of mouse serum inflammatory factor determination

[0093] The sampling results on the 9th day showed that compared with the blank group, the IL-10 level in the model group was significantly decreased (P < 0.05), and the TNF-α level was significantly increased (P < 0.001), indicating that the modeling was effective; compared with the model group, the IL-6 level in the positive drug gabapentin group and the experimental drug HEO medium and high dose groups was significantly decreased (P < 0.01); the TNF-α level in the HEO medium and high dose groups was significantly decreased (P < 0.01); the MCP-1 level in the gabapentin group and the HEO low and high dose groups was significantly decreased (P < 0.05); and compared with the HEO group, the MCP-1 in the group that was given the CB2 antagonist AM630 at the same time was significantly decreased (P < 0.05).

[0094] The sampling results on the 15th day showed that there was no significant difference in the levels of inflammatory factors among the groups.

[0095] For specific results, see Figure 4 , Figure 5 , Figure 6 , Figure 7 And Table 6, Table 7.

[0096] Table 6 Mouse IL-6, IL-10 determination results ( Day 9 n=4 and day 15 n=6)

[0097]

[0098] Note: Compared with the blank control group: #-P<0.05, ###-P<0.001; Compared with the model group: *-P<0.05; **-P<0.01; ***-P<0.001; Compared with the HEO-M group: @ -P<0.05.

[0099] Table 7 Mouse TNF-α, MCP-1 determination results ( Day 9 n=4 and day 15 n=6)

[0100]

[0101]

[0102] Note: Compared with the blank control group: #-P<0.05, ###-P<0.001; Compared with the model group: *-P<0.05; **-P<0.01; ***-P<0.001; Compared with the HEO-M group: @ -P<0.05.

Claims

1. A pharmaceutical composition, characterized in that It contains: β-caryophyllene, α-humulene and caryophyllene oxide.

2. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition also includes one or more of (-)-humulene epoxide II, α-bisabolol and (+)-delta-cadinene, cnidium-3,7(11)-diene, γ-ylang-ylangolene, hordein, β-serene, α-bergamotene and bergamotene oxide.

3. The pharmaceutical composition according to claim 2, characterized in that The pharmaceutical composition satisfies one or more of the following: (1) The β-caryophyllene content is preferably 10-35%; more preferably 15-30%; for example 26.49%; (2) The α-humulene content is preferably 5 to 20%; more preferably 10 to 15%; for example 12.11%; (3) The content of caryophyllene oxide is preferably 8-20%; more preferably 10-15%; for example 11.65%; (4) The content of (-)-humulene epoxide II is preferably 3 to 5%, for example 3.76%. (5) The content of α-bisabolol is preferably 2-5%, for example 3.27%. (6) The (+)-δ-cadinene content is preferably 2-5%, for example 3.24%. (7) The content of cnidium-3,7(11)-diene is preferably 2 to 5%, for example 3.07%. (8) The γ-ylangolene content is preferably 2-4%, for example 2.91%. (9) The hordein content is preferably 2-4%, for example 2.83%. (10) The β-serene content is preferably 2-4%, for example 2.44%. (11) The α-bergamotene content is preferably 2-4%, for example 2.37%. (12) The content of bergamot oxide is preferably 1-3%, for example 1.88%.

4. The pharmaceutical composition according to claim 3, characterized in that The pharmaceutical composition satisfies one or more of the following: (1) The active ingredients of the pharmaceutical composition are derived from plant essential oils; (2) The plant is industrial hemp flowers and leaves; (3) The active ingredient of the pharmaceutical composition is a sesquiterpenoid compound.

5. The pharmaceutical composition according to claim 4, characterized in that The active ingredients of the pharmaceutical composition consist of active substances beta-caryophyllene, alpha-humulene and caryophyllene oxide.

6. The pharmaceutical composition according to claim 4, characterized in that The active ingredients of the pharmaceutical composition are β-caryophyllene, α-humulene and caryophyllene oxide, (-)-humulene epoxide II, α-bisabolol and (+)-delta-cadinene, cnidium-3,7(11)-diene, γ-ylang-ylangolene, hordein, β-thulene, α-bergamotene and bergamotene oxide.

7. The pharmaceutical composition according to any one of claims 1 to 6, characterized in that The pharmaceutical composition is prepared by the following method, which comprises the following steps: (1) extracting and collecting volatile oil from industrial hemp flower and leaf powder by steam distillation; (2) subjecting the collected volatile oil to gas phase separation to obtain the pharmaceutical composition according to any one of claims 1 to 6; Preferably, the solvent of the steam distillation method is water; And / or, the distillation temperature of the steam distillation method is 150°C to 170°C; for example, 160°C; And / or, the gas phase chromatographic column is a HP-5MSUI column; And / or, the gas phase injection port temperature is 200°C to 280°C; for example, 250°C; And / or, the gas phase chromatographic column flow rate is 0.8 ml / min to 12 ml / min; for example, 1 ml / min; And / or, the gas phase chromatographic column split ratio is 1:1 to 3:1; for example, 2:1; More preferably, the gas phase is programmed to heat, and the method of programmed heating is preferably: initial temperature 60°C, maintained for 2 minutes; increased to 110°C at 20°C / min; then increased to 160°C at 2°C / min, maintained for 1 minute; and then increased to 280°C at 20°C / min, maintained for 3 minutes.

8. Use of the pharmaceutical composition according to any of claims 1 to 7 in the preparation of a medicament for preventing and / or treating peripheral neuropathy pain; Preferably, the drug is used to increase the activity of SOD; and / or, the drug is used to alleviate oxidative stress; and / or, the drug is used to reduce sensitivity to thermal stimulation and / or sensitivity to mechanical stimulation; And / or, the medicament is used for preventing and / or treating pain caused by inflammation.

9. The use according to claim 8, wherein the peripheral neuropathy pain is mediated by one or more inflammatory cytokines selected from the group consisting of IL-6, IL-10, TNF-α and MCP-1.

10. A pharmaceutical composition as claimed in claims 1-7, wherein the pharmaceutical composition is used in the preparation of an inhibitor for inhibiting the secretion of inflammatory cytokines, preferably, the inhibitor is used to inhibit the secretion of one or more inflammatory cytokines among IL-6, IL-10, TNF-α and MCP-1.

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