Extraction method of safflower leaf exosome-like nano-vesicles and application of safflower leaf exosome-like nano-vesicles in preparation of medicine for preventing and treating adriamycin-induced cardiotoxicity

Abstract

The invention discloses an extraction method of safflower leaf exosome-like nano-vesicles and application of the safflower leaf exosome-like nano-vesicles in preparation of drugs for preventing and treating adriamycin-induced cardiotoxicity, fresh safflower leaves are taken and cleaned, a PBS solution is added for crushing and filtering, filtrate is collected and continuously centrifuged, supernate is taken and subjected to ultracentrifugation, precipitates are collected after centrifugation, and the safflower leaf exosome-like nano-vesicles are obtained; and filtering and sterilizing by using a filter membrane to obtain the safflower leaf exosome-like nano-vesicles. The carthamus tinctorius leaf exosome-like nano-vesicles can significantly improve the heart function of doxorubicin-induced cardiotoxic mice; myocardial injury induced by doxorubicin can be repaired, and myocardial fibrosis induced by doxorubicin can be inhibited; the content of a myocardial injury marker lactic dehydrogenase in body serum can be reduced, a new treatment strategy is provided for preventing and treating adriamycin-induced cardiotoxic diseases, and the application prospect is good.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the extraction method of exosome-like nanovesicles from safflower leaves and their application in the preparation of drugs for preventing and treating doxorubicin-induced cardiotoxicity. Background Technology

[0002] Doxorubicin (DOX) is an anthracycline compound widely used in chemotherapy for various malignant tumors, including breast cancer, lung cancer, and lymphoma. As a chemotherapeutic drug, DOX kills tumor cells but also causes toxicity to normal cardiomyocytes, leading to cardiotoxicity, a major side effect. Doxorubicin-induced cardiotoxicity (DIC) mainly manifests as cardiomyocyte apoptosis, myocardial dysfunction, and heart failure. This toxicity can develop in a subacute, acute, or chronic manner, and risk factors include cumulative dose, drug administration rate, pre-existing heart disease, and hypertension. Studies have found that the risk of death from drug-related cardiotoxicity in cancer patients exceeds the risk of death from the tumor itself or from tumor recurrence. Therefore, developing effective and safe drugs for the prevention and treatment of DIC is currently a key research focus.

[0003] safflower Carthamus tinctorius Safflower (L.), also known as safflower or thorny safflower, is a plant belonging to the genus Safflower in the family Asteraceae. Its dried tubular flowers are orange-red with narrow tubes and a distinctive fragrance. It is a traditional and precious Chinese medicinal herb and one of the most widely used herbs in Mongolian medicine. As an important medicinal plant, safflower is mainly produced in Henan, Hunan, Sichuan, Xinjiang, and Tibet, and has the effects of promoting blood circulation, removing blood stasis, dispelling dampness, and reducing swelling. Historically, the use and research of safflower have mainly focused on its flowers. However, the low flower yield of safflower plants greatly limits its research and development value. After the flowers are harvested for medicinal purposes, a large amount of safflower leaves are discarded, resulting in resource waste. However, safflower leaves also have significant application value. Safflower leaves, a plant used both as food and medicine, are recorded in the 5,000-year-old Ayurvedic medical tradition as having laxative, appetite-stimulating, and diuretic effects, and can be used to relieve urethritis and eye diseases. Modern research has shown that, in addition to alkaloids and terpenes found in safflower flowers, safflower leaves also contain flavonoids and polyphenols, exhibiting excellent antioxidant and free radical scavenging capabilities. Beyond its medicinal potential, safflower leaves are commonly consumed as a vegetable in China, India, Pakistan, and Myanmar by blanching the tender leaves. After drying, they can also be made into functional leaf teas, becoming an important source of health-promoting components in the daily diet.

[0004] Exosome-like vesicles (also known as cellular vesicles or extracellular vesicles) are natural lipid bilayer nanoparticles capable of carrying various biomolecules, including proteins, lipids, DNA, and RNA, which is important for intercellular communication and regulation. Plant-derived exosome-like nanovesicles are safer, less toxic, and less immunogenic than animal-derived exosomes. In recent years, medicinal plant-derived exosome-like nanovesicles have shown good therapeutic relevance in antioxidation and anti-inflammation, and have also demonstrated broad application potential in the medical and cosmetic fields. For example, patent document CN120643477A discloses that Himalayan Mirabilis exosome vesicles can scavenge free radical ions and inhibit the production of reactive oxygen species in cells, and can be used to prepare whitening and antioxidant products; patent document CN120361090A discloses that Polygonum multiflorum exosome vesicles can scavenge DPPH free radicals, ABTS free radicals, and hydroxyl free radicals, and have good antioxidant activity, while inhibiting hyaluronidase activity, and have good anti-inflammatory activity; extracellular vesicles derived from Saposhnikovia divaricata (CN120459158A), extracellular vesicles derived from Houttuynia cordata (CN118593592A), extracellular vesicle nanoparticles derived from Atractylodes macrocephala (J Nanobiotechnology. 2025, 23(1):433), and exosome vesicles derived from Coix lacryma-jobi seeds (Int J Biol Macromol. 2025, Both 332:148799) have a certain therapeutic effect on ulcerative colitis and can reduce the expression of inflammatory factors in macrophages; exosomes and vesicles derived from bitter melon can reduce the accumulation of reactive oxygen species and have a protective effect against cardiac damage caused by doxorubicin (J Nanobiotechnology.2024, 22(1):464).

[0005] However, there is almost no research on exosome-like nanovesicles from safflower leaves in the current technology, and there are no reports on their ability to alleviate doxorubicin-induced cardiotoxicity. Therefore, developing a method for extracting and preparing exosomes from safflower leaves and applying the prepared safflower leaf exosomes to the preparation of drug products for preventing and treating doxorubicin-induced cardiotoxicity is of great significance and promise. Summary of the Invention

[0006] The purpose of this invention is to provide a method for extracting exosome-like nanovesicles from *Carthamus tinctorius* leaves and its application in the preparation of drugs for preventing and treating doxorubicin-induced cardiotoxicity. The above extraction method can extract high-purity exosome-like nanovesicles from *Carthamus tinctorius* leaves with a high extraction rate, simple operation, and low cost. The exosome-like nanovesicles extracted by this invention can significantly reduce doxorubicin-induced cardiotoxicity and have a significant effect on improving cardiac function.

[0007] To achieve the above objectives, the present invention employs the following technical solution: a method for extracting exosome-like nanovesicles from safflower leaves. The specific extraction steps are as follows: fresh safflower leaves are cleaned, added to phosphate-buffered saline (PBS) and ground or juiced to form a homogenate. The homogenate is filtered through gauze to collect the filtrate. The filtrate is then centrifuged at differential speed to collect the supernatant. Finally, the precipitate is collected by ultra-high speed centrifugation to obtain safflower leaf exosome-like nanovesicles. The safflower leaf exosome-like nanovesicles are spherical or saucer-shaped, have a double membrane structure, a particle size range of 48.0~198.0 nm, an average particle size of 105.7 nm, and a zeta potential of -17.7 mV.

[0008] Furthermore, the differential centrifugation process is as follows: the filtrate is centrifuged at 2-8℃ and 100-3000g for 10-20 min, and the supernatant is collected; the collected supernatant is centrifuged at 2-8℃ and 3000-8000g for 20-40 min, and the supernatant is collected; the collected supernatant is centrifuged at 2-8℃ and 8000-12000g for 40-80 min, and the supernatant is collected; the supernatant is filtered using a sterile vacuum filtration flask with a pore size of 0.22μm to remove bacteria, and the collected filtrate is centrifuged at 2-8℃ and 50000-200000g for 60-180 min, the supernatant is discarded, and the precipitate is resuspended using PBS; then, the filtrate is filtered through a filter membrane to obtain red flower leaf exosome-like nanovesicles.

[0009] The application of the safflower leaf exosome-like nanovesicles described in this invention in the preparation of drugs for preventing and treating doxorubicin-induced cardiotoxicity.

[0010] Furthermore, the safflower leaf exosome-like nanovesicles are used to improve cardiac function affected by mycotoxin-induced cardiotoxicity.

[0011] Furthermore, the safflower leaf exosome-like nanovesicles are used to repair doxorubicin-induced myocardial damage.

[0012] Furthermore, the safflower leaf exosome-like nanovesicles are used to inhibit doxorubicin-induced myocardial fibrosis.

[0013] Furthermore, the safflower leaf exosome-like nanovesicles are used to reduce the level of lactate dehydrogenase, a marker of myocardial injury, in the body's serum.

[0014] A drug for the prevention and / or treatment of doxorubicin-induced cardiotoxicity, said drug comprising an effective amount of safflower exosome-like nanovesicles and a pharmaceutically acceptable carrier.

[0015] Furthermore, the concentration of safflower leaf exosome-like nanovesicles in the drug is ≤120 μg / mL.

[0016] Furthermore, the dosage form of the drug includes tablets, capsules, pills, powders, granules, suspensions, oral solutions, powder for injection, or injections.

[0017] This invention has the following advantages and beneficial effects: The extraction method of this invention can extract high-purity safflower leaf exosome-like nanovesicles with a high extraction rate, is simple to operate, low in cost, and suitable for industrial application. The safflower leaf exosome-like nanovesicles extracted by this invention can significantly improve cardiac function in mice with doxorubicin-induced cardiotoxicity; can repair doxorubicin-induced myocardial damage and inhibit doxorubicin-induced myocardial fibrosis; and can reduce the level of lactate dehydrogenase, a marker of myocardial injury, in serum, providing a new treatment strategy for the prevention and treatment of doxorubicin-induced cardiotoxicity, and has good application prospects. Attached Figure Description

[0018] Figure 1 The characterization results of the exosome-like nanovesicles extracted from red peony leaves in Example 1 are shown in the scanning electron microscope image, particle size distribution map, and potential map.

[0019] Figure 2 The image shows the percentage of cell viability of rat cardiomyocytes H9C2 cells extracted from Carthamus tinctorius leaf-derived nanovesicles (CDNVs) in Example 1, as well as the uptake of fluorescently labeled CDNVs by rat cardiomyocytes H9C2 cells at different time points.

[0020] Figure 3 Representative images and statistical results of cardiac function ultrasound in mice of each group are shown. Control represents the blank control group (mice that received only saline injection and did not receive doxorubicin treatment); CDNVs represent the control group treated only with safflower leaf exosome-like nanovesicles; DOX represents the doxorubicin-induced injury model group; and DOX+CDNVs represent the group treated with both doxorubicin induction and safflower leaf exosome-like nanovesicles.

[0021] Figure 4 HE staining images and Masson staining images of the heart morphology of mice in each group, as well as the content of cardiac injury markers in the serum of mice in each group, are shown. Control represents the blank control group (mice that received only saline injection and did not receive doxorubicin treatment); CDNVs represents the control group treated only with safflower leaf exosome-like nanovesicles; DOX represents the doxorubicin-induced injury model group; DOX+CDNVs represents the group treated with doxorubicin induction and safflower leaf exosome-like nanovesicles.

[0022] Note: Compared with the model group, *P<0.05, **P<0.01, ***P<0.001, n=6. Detailed Implementation

[0023] The following examples further illustrate the above-described content of the present invention in detail, but should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Unless otherwise specified, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0024] Firstly, this invention provides a method for extracting exosome-like nanovesicles from *Carthamus tinctorius* leaves, mainly by the following steps: Fresh *Carthamus tinctorius* leaves are washed and dried, mixed with a pre-cooled PBS solution (pH=7.4) at 4°C, and ground in a mortar or juiced. The resulting juice is filtered twice with a filter cloth to remove residue. Then, the mixture is continuously centrifuged at differential speed at 4°C. The supernatant is collected after centrifugation and further subjected to ultracentrifugation. The precipitate is collected and filtered through a filter membrane for sterilization, thus obtaining the exosome-like nanovesicles from *Carthamus tinctorius* leaves. The above differential centrifugation method is simple and convenient, and can effectively extract exosome-like nanoparticles from *Carthamus tinctorius* leaves. The crushing method includes, but is not limited to, grinding or juicing.

[0025] Secondly, this invention provides the application of the aforementioned *Carthamus tinctorius* exosome-like nanovesicles in the prevention or treatment of doxorubicin-induced cardiotoxicity. The *Carthamus tinctorius* exosome-like nanoparticles provided by this invention significantly reduce doxorubicin-induced cardiotoxicity, repair doxorubicin-induced myocardial damage, inhibit doxorubicin-induced myocardial fibrosis, and have a significant effect on improving cardiac function. Example 1

[0026] Preparation of exosome-like nanovesicles from safflower leaves: Wash and dry the safflower leaves, mix them with pre-cooled phosphate buffer (pH=7.4) at 4℃, and juice them using a screw press. Filter the resulting juice twice through a filter cloth to remove residue and collect the filtrate. Process the filtrate using differential centrifugation. First, centrifuge at 1000×g, 4℃ for 20 min to remove larger particles from the safflower leaves; discard the precipitate. Centrifuge the collected supernatant at 3000×g, 4℃ for 20 min to remove plant fibers from the safflower leaves; discard the precipitate again. Centrifuge the collected supernatant at 10000×g... Centrifuge at 100000×g, 4℃ for 60 min to remove small particles; discard the precipitate, collect the supernatant, and filter the supernatant through a sterile 0.22μm filtration flask for sterilization. Finally, centrifuge the supernatant at 100000×g, 4℃ for 90 min, collect the precipitate, resuspend the precipitate in pre-cooled PBS, and then filter again through a 0.22μm sterile filter membrane to obtain a *Caragana korshinskii* exosome-like nanovesicle solution. Quantify the protein content of the collected *Caragana korshinskii* exosome-like nanovesicles according to the BCA protein quantification kit method, and store at -80℃ for later use. All subsequent experiments were conducted using the *Caragana korshinskii* exosome-like nanovesicles obtained in Example 1.

[0027] Characterization of red flower leaf exosome-like nanovesicles: Particle size distribution and potential were determined using Malvern Zetasizer Nano ZS microscopy; morphology of safflower leaf exosome-like nanovesicles was observed using TEM transmission electron microscopy. TEM electron microscopy, particle size distribution, and potential of safflower-derived exosome-like nanovesicles are shown below. Figure 1 As shown in A, B, and C, TEM revealed that the isolated red peony leaf exosome-like nanovesicles had a typical exosome vesicle structure, with particle sizes ranging from 48.0 nm to 198.0 nm and an average particle size of 105.7 nm. TEM electron microscopy also showed that the particle sizes ranged from 50 to 200 nm, and the zeta potential of the red peony leaf exosome-like nanovesicles was around -17.7 mV. Example 2

[0028] The activity of *Caragana korshinskii* exosome-like nanovesicles on H9C2 rat cardiomyocytes and the uptake of *Caragana korshinskii* exosome-like nanoparticles by H9C2 cells were evaluated. Cell viability was assessed using a CCK-8 assay kit. The test results are as follows: Figure 2 As shown in Figure A, it can be seen from the figure that red peony leaf exosome-like nanovesicles at concentrations not exceeding 120 μg / mL have no toxicity to cells.

[0029] To further verify the protective effect of *Caragana korshinskii* exosome-like nanovesicles on doxorubicin-treated H9C2 cells, the Control group consisted of normally cultured H9C2 cells, the Dox group consisted of H9C2 cells treated with 0.25 μM doxorubicin, the CDNVs group consisted of H9C2 cells treated with only 30 μg / mL *Caragana korshinskii* exosome-like nanovesicles, and the Dox+CDNVs group consisted of H9C2 cells treated with both 0.25 μM doxorubicin and 30 μg / mL *Caragana korshinskii* exosome-like nanovesicles. The results are as follows: Figure 2 As shown in Figure B, it can be seen that 30 μg / mL of safflower leaf exosome-like nanovesicles can significantly improve the survival rate of cardiomyocytes damaged by doxorubicin.

[0030] Red clover leaf exosome-like nanoparticles labeled with the red fluorescent dye Dil were co-incubated with H9C2 cells for 6 h, 12 h, and 24 h, respectively. Their uptake was observed by the intensity of the red fluorescence. Figure 2 As shown in Figure C, it can be seen that the red peony leaf exosome-like nanovesicles can be taken up by H9C2 cells, and the amount of red peony leaf exosome-like nanovesicles taken up increases with time. Example 3

[0031] (1) Constructing a mouse model of doxorubicin-induced cardiotoxicity Doxorubicin-induced cardiotoxicity model was established using 6-8 week old male C57BL / 6 mice. The specific steps were as follows: After 2 weeks of acclimatization, the mice were randomly divided into a blank control group (Control), a red peony leaf exosome-like nanovesicle control group (CDNVs), a doxorubicin model group (Dox), and a doxorubicin and red peony leaf exosome-like nanoparticle administration group (Dox+CDNVs), with 6 mice in each group.

[0032] The blank control group and the redbud exosome-like nanovesicle control group were injected with sterile saline via tail vein. The doxorubicin group and the doxorubicin + redbud exosome-like nanovesicle administration group were injected with 5 mg / kg of doxorubicin via tail vein, once daily for four days starting from day 1 (cumulative dose 20 mg / kg) to establish a cardiotoxicity model. During administration, the blank control group and the doxorubicin model group were injected with sterile PBS via tail vein as controls. The redbud exosome-like nanoparticle control group and the doxorubicin + redbud exosome-like nanovesicle administration group were injected with redbud exosome-like nanovesicles (40 mg / kg) via tail vein, administered on day 0 and day 3, twice a day (cumulative dose 80 mg / kg). The efficacy of the administration was assessed on day 6.

[0033] (2) Echocardiography in mice Mice were anesthetized, and their left chest was prepared. They were then placed on an ultrasound worktable with their limbs pressed against copper plates and coupling gel applied. The Vevo2100 small animal ultrasound imaging system was used to measure the left ventricular end-diastolic diameter and left ventricular end-systolic diameter. Ejection fraction (EF) and fractional shortening (FS) values ​​were calculated using ultrasound data analysis software, thus assessing cardiac function.

[0034] Experimental results: Compared with the control group, the left ventricular ejection fraction (EF) and left ventricular shortening fraction (FS) of mice in the doxorubicin group decreased (P<0.01), indicating that doxorubicin injection caused cardiac function impairment in mice.

[0035] Compared with the model group, mice in the safflower leaf exosome-like nanovesicle (80 mg / kg) treatment group showed significantly increased EF and FS values ​​(P<0.01), indicating that safflower leaf exosome-like nanovesicles can significantly alleviate doxorubicin-induced cardiac function decline (e.g., Figure 3 (As shown).

[0036] (3) HE staining to observe pathological changes in mouse heart tissue Heart tissue was immersed in 4 wt% paraformaldehyde solution for 24 hours, then routinely embedded in paraffin. After dewaxing, sections were stained with hematoxylin for 2 minutes, rinsed with running water for 5 minutes, stained with eosin for 90 seconds, rinsed with running water for 10 minutes, dried at 60℃ for 30 minutes, mounted with neutral resin, and the pathological changes of heart tissue in each group were observed under a microscope.

[0037] Experimental results: In the control group, the cardiomyocytes of mice were morphologically intact, neatly arranged, and uniformly stained, with no obvious structural changes in the nuclei and no inflammatory cell infiltration. In contrast, the cardiomyocytes of mice in the doxorubicin group showed a clearly loose and disordered arrangement, with indistinct or even absent striations on the myocardial fibers, pyknosis or rupture of the nuclei, and inflammatory cell infiltration in the myocardial interstitium, resulting in loss of normal structure. Compared to the doxorubicin group, the arrangement of myocardial tissue and the dissolution of myocardial fibers were improved in mice with safflower leaf exosome-like nanovesicles; only a small number of cells lost their normal structure, and occasional inflammatory cell infiltration was observed. Figure 4 As shown in Figure A. The results indicate that safflower leaf exosome-like nanovesicles can repair doxorubicin-induced myocardial injury.

[0038] (4) Masson staining Dewax paraffin sections were prepared and stained with hematoxylin for 5-10 min. Differentiation was then performed in 1 wt% hydrochloric acid-alcohol solution for 5-15 s, followed by rinsing with water. The sections were then kept in 0.1 wt%-1 wt% lithium carbonate solution for 5 min to increase the inversion intensity, rinsed with water, and stained with Ponceau S acidic fuchsin for 5-10 min. Afterward, the sections were rinsed with a weak acid working solution (2 wt% glacial acetic acid aqueous solution) for 1 min. Differentiation was then performed in 1 wt% phosphomolybdic acid aqueous solution for 3-5 min, followed by rinsing with a weak acid working solution for 1 min. Aniline blue solution was stained for 1-2 min, followed by rinsing with a weak acid working solution for 1 min. The sections were then dehydrated, mounted, and observed under a microscope.

[0039] Experimental results: In the control group, the cardiomyocytes of mice were dense, well-organized, and showed no obvious collagen deposition. In the doxorubicin group, the cardiomyocytes of mice were disordered and loosened, with significant collagen deposition forming scars, and the degree of fibrosis was significantly aggravated. In the group treated with safflower leaf exosome-like nanovesicles (80 mg / kg), collagen deposition in the cardiomyocytes of mice was significantly reduced. These results indicate that safflower leaf exosome-like nanovesicles can reduce collagen deposition in myocardial tissue and alleviate the degree of myocardial fibrosis. Figure 4 As shown in B and C.

[0040] Furthermore, compared with the doxorubicin group, the level of lactate dehydrogenase (LDH), a marker of myocardial injury, was significantly reduced in the serum of mice in the safflower leaf exosome-like nanovesicle group (80 mg / kg). Figure 4 As shown in D.

[0041] The foregoing has shown and described the basic principles, main features and advantages of the present invention. Various changes and modifications can be made to the present invention without departing from the spirit and scope thereof, and all such changes and modifications fall within the scope of the present invention as claimed.

Claims

1. A method for extracting safflower leaf exosome-like nanovesicles, characterized by The specific extraction steps are as follows: the fresh safflower leaves are washed clean, phosphate buffer is added for grinding or juicing to break into a homogenate, the homogenate is filtered through gauze to collect the filtrate, the filtrate is subjected to differential centrifugation to take the supernatant, and finally, the supernatant is subjected to ultrahigh-speed centrifugation to collect the precipitate to obtain safflower leaf exosome-like nanovesicles, wherein the safflower leaf exosome-like nanovesicles are circular or saucer-shaped, have a double-membrane structure, the particle size interval is 48.0-198.0 nm, the average particle size is 105.7 nm, and the Zeta potential is-17.7 mV.

2. The method of claim 1, wherein the safflower leaf exosome-like nanovesicles are extracted from the safflower leaves. The differential centrifugation process is as follows: the filtrate is centrifuged at 2-8 DEG C and 100-3000 g for 10-20 min to collect the supernatant; the collected supernatant is centrifuged at 2-8 DEG C and 3000-8000 g for 20-40 min to collect the supernatant; and the collected supernatant is centrifuged at 2-8 DEG C and 8000-12000 g for 40-80 min to collect the supernatant. The supernatant is filtered through a sterile filter bottle with a pore size of 0.22 μm to remove bacteria, and the collected filtrate is centrifuged at 2-8 DEG C and 50000-200000 g for 60-180 min, the supernatant is discarded, and the precipitate is resuspended with PBS; then the filtrate is filtered through a filter membrane to obtain safflower leaf exosome-like nanovesicles.

3. The safflower leaf exosome-like nanovesicles extracted by the method of any one of claims 1-2 are used for preparing a drug for preventing and treating adriamycin-induced cardiotoxicity.

4. Use according to claim 3, characterized in that: The safflower leaf exosome-like nanovesicles are used for improving the heart function of adriamycin-induced cardiotoxicity.

5. Use according to claim 3, characterized in that: The safflower leaf exosome-like nanovesicles are used for repairing adriamycin-induced myocardial injury.

6. Use according to claim 3, characterized in that: The safflower leaf exosome-like nanovesicles are used for inhibiting adriamycin-induced myocardial fibrosis.

7. Use according to claim 3, characterized in that: The safflower leaf exosome-like nanovesicles are used for reducing the content of serum myocardial injury marker lactate dehydrogenase in the body.

8. A medicament for preventing and / or treating doxorubicin-induced cardiotoxicity disease, characterized by: The drug comprises an effective amount of the safflower leaf exosome-like nanovesicles extracted by the method of any one of claims 1-2 and a pharmaceutically acceptable carrier.

9. The medicament according to claim 8, characterized in that: The concentration of the safflower leaf exosome-like nanovesicles in the drug is ≤120 μg / mL.

10. The medicament according to claim 8, characterized in that: The dosage form of the drug includes tablets, capsules, pills, powders, granules, suspensions, oral solutions, powder injections or injections.

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