Use of dacarbazine in the preparation of inhibitors of streptococcus mutans

Abstract

The application discloses application of dacarbazine in preparation of an inhibitor of streptococcus mutans and belongs to the technical field of biological medicines. Researches show that the dacarbazine has a significant inhibitory effect on the activity and growth of the streptococcus mutans, can significantly down-regulate the expression of gtfB, gtfC, gtfD, gbpC and / or srtA genes of the streptococcus mutans, and effectively inhibit the formation and activity of a biofilm of the streptococcus mutans. The application provides a new drug selection for the prevention and treatment of the streptococcus mutans and has a wide application prospect in the prevention, improvement and treatment of dental caries in the clinic.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the application of dacarbazin in the preparation of inhibitors of Streptococcus mutans. Background Technology

[0002] Dental caries is a common chronic oral disease. World Health Organization (WHO) data on oral health shows that a large number of people worldwide are affected by oral diseases, and untreated permanent tooth caries remains one of the most burdensome oral health problems. The occurrence of dental caries is closely related to the colonization of cariogenic bacteria in the tooth surface biofilm, acid production from sugar metabolism, and demineralization of the hard tooth tissue. Streptococcus mutans (Streptococcus mutans) Streptococcus mutans (Illicium verum) is a facultative anaerobic Gram-positive bacterium and one of the important caries-related microorganisms in the process of caries development. This bacterium can adhere to the enamel surface and form a biofilm in the presence of sucrose. The bacteria in the biofilm continuously produce acid through metabolism, which can lead to a decrease in local pH, promote demineralization of the hard tissue of the tooth and form caries lesions.

[0003] The formation of biofilms and caries-related phenotypes in Streptococcus mutans are regulated by multiple genes, among which gtfB, gtfC, gtfD, srtA, and gbpC are associated with extracellular dextran synthesis, bacterial adhesion, and biofilm stability. Therefore, inhibiting the growth, biofilm formation, and acid production of Streptococcus mutans is an important intervention direction in caries prevention and control technologies.

[0004] Currently, dental caries prevention and control mainly relies on measures such as mechanical plaque removal, topical fluoride application, and oral antibacterial agents. These methods have clear value in reducing the risk of dental caries, but are still limited by factors such as adherence to treatment, duration of application, plaque recolonization, and the impact on the oral microecology. Therefore, screening for novel anti-caries agents is of great significance. Summary of the Invention

[0005] To overcome the aforementioned defects and shortcomings in the prior art, the present invention provides the application of dacarbazin in the preparation of inhibitors of Streptococcus mutans.

[0006] The first objective of this invention is to provide the use of dacarbazin in the preparation of inhibitors of Streptococcus mutans.

[0007] A second objective of this invention is to provide the use of dacarbazin in the preparation of inhibitors for biofilm formation of Streptococcus mutans.

[0008] A third object of the present invention is to provide the use of dacarbazin in the preparation of medicaments for the prevention and / or treatment of Streptococcus mutans infections.

[0009] A fourth objective of this invention is to provide the use of dacarbazin in the preparation of medicaments for inhibiting the expression of the gtfB, gtfC, gtfD, gbpC and / or srtA genes in Streptococcus mutans.

[0010] A fifth object of the present invention is to provide the use of dacarbazin in the preparation of medicaments for the prevention, improvement and / or treatment of diseases caused by Streptococcus mutans.

[0011] A sixth object of the present invention is to provide the use of dacarbazin in the preparation of medicaments for the prevention, improvement and / or treatment of dental caries.

[0012] This invention claims protection for the following: Application of dacarbazin in the preparation of inhibitors of Streptococcus mutans.

[0013] Application of dacarbazin in the preparation of inhibitors for biofilm formation of Streptococcus mutans.

[0014] The use of dacarbazin in the preparation of drugs for the prevention and / or treatment of Streptococcus mutans infections.

[0015] Application of dacarbazin in the preparation of drugs that inhibit the expression of gtfB, gtfC, gtfD, gbpC and / or srtA genes in Streptococcus mutans.

[0016] The use of dacarbazin in the preparation of medicines for the prevention, improvement and / or treatment of diseases caused by Streptococcus mutans.

[0017] The use of dacarbazine in the preparation of medicines for the prevention, improvement and / or treatment of dental caries.

[0018] Preferably, the drug inhibits the growth of Streptococcus mutans and / or inhibits the formation of Streptococcus mutans biofilm.

[0019] Streptococcus mutans ATCC 700710 is a pathogenic bacterium that is highly adapted to the oral environment and has high prevalence and high ecological adaptability. Preferably, the inhibition of Streptococcus mutans is the inhibition of Streptococcus mutans ATCC 700610.

[0020] Preferably, the effective concentration of dacarbazine is 3–70 μg / mL.

[0021] More preferably, the effective concentration of dacarbazine is 4–64 μg / mL.

[0022] More preferably, the effective concentration of dacarbazine is 16–64 μg / mL.

[0023] As one possible approach, the drug may also include pharmaceutically acceptable excipients.

[0024] As one possible approach, the dosage form of the drug includes tablets, capsules, granules, powders, and pills.

[0025] Compared with the prior art, the present invention has the following beneficial effects: This invention discloses the application of dacarbazin in the preparation of inhibitors against Streptococcus mutans. The research of this invention shows that dacarbazin has a significant inhibitory effect on the activity and growth of Streptococcus mutans, and can significantly downregulate the expression of Streptococcus mutans gtfB, gtfC, gtfD, gbpC and / or srtA genes, and effectively inhibit the formation and activity of Streptococcus mutans biofilm. This invention provides a new drug option for the prevention and treatment of Streptococcus mutans, and has broad application prospects in the clinical prevention, improvement and treatment of dental caries. Attached Figure Description

[0026] Figure 1 The results of the MIC experiment of dacarbazin against Streptococcus mutans in Example 1 are shown.

[0027] Figure 2 The 24-hour growth curve of Streptococcus mutans under dacarbazin treatment.

[0028] Figure 3 Crystal violet staining results of Streptococcus mutans biofilm after treatment with different concentrations of dacarbazine.

[0029] Figure 4 Quantitative results of biomass of Streptococcus mutans biofilm after treatment with different concentrations of dacarbazine.

[0030] Figure 5 The results of live / dead staining of Streptococcus mutans biofilm after treatment with different concentrations of dacarbazine.

[0031] Figure 6 Scanning electron microscope images of Streptococcus mutans biofilms after treatment with different concentrations of dacarbazine.

[0032] Figure 7 MTT staining results of mature biofilms after treatment with different concentrations of dacarbazine.

[0033] Figure 8 Quantitative results of MTT absorbance of mature biofilms after treatment with different concentrations of dacarbazine.

[0034] Figure 9 The amount of lactic acid produced by the biofilm of Streptococcus mutans after treatment with different concentrations of dacarbazine.

[0035] Figure 10 The pH changes of Streptococcus mutans culture systems treated with different concentrations of dacarbazine within 24 h.

[0036] Figure 11The effect of dacarbazin treatment on the expression levels of gtfB, gtfC, gtfD, gbpC and srtA in Streptococcus mutans.

[0037] Figure 12 Micro-CT images of rat molars from different treatment groups.

[0038] Figure 13 The results of ammonium urea staining of rat molars in different treatment groups.

[0039] Figure 14 The Keyes scores of rat molars in different treatment groups were statistically analyzed.

[0040] Figure 15 The results of H&E staining of buccal mucosa, gingiva, tongue, liver, spleen and kidney of rats in different treatment groups. Detailed Implementation

[0041] The present invention will be further illustrated below with reference to specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field.

[0042] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0043] The Streptococcus mutans used in the examples was Streptococcus mutans ATCC 700610.

[0044] Example 1: MIC value of dacarbazin against Streptococcus mutans I. Experimental Methods Streptococcus mutans stored at -80℃ was streaked onto BHI agar plates and anaerobically cultured at 37℃ for 24 h. Single colonies were then selected and inoculated onto BHI broth medium and anaerobically cultured at 37℃ for 24 h. The bacterial suspension was then resuspended in BHI broth medium, and the final inoculum was adjusted to 1×10⁻⁶. 6 CFU / mL. Bacterial suspension and serially diluted dacarbazin solution of equal volume were added to 96-well plates to achieve final concentrations of dacarbazin of 2048, 1024, 512, 256, 128, 64, 32, 16, 8, 4, and 2 μg / mL, respectively. The control group received no dacarbazin solution, while the blank group received only BHI broth medium. After 24 h of incubation, the lowest drug concentration at which no obvious precipitation or turbidity was observed was determined as the MIC.

[0045] II. Experimental Results The results are as follows Figure 1 As shown, no visible bacterial precipitate was observed in the treatment group with a final dacarbazin concentration of 8 μg / mL, indicating that the MIC value of dacarbazin against Streptococcus mutans is 8 μg / mL.

[0046] Example 2: Inhibitory effect of dacarbazin on the in vitro growth of Streptococcus mutans I. Experimental Methods Overnight Streptococcus mutans culture was resuspended in fresh BHI broth, and dacarbazin solution diluted serially with BHI broth was added to achieve final concentrations of 4, 16, and 64 μg / mL (corresponding to 1 / 2×MIC, 2×MIC, and 8×MIC, respectively). A solvent growth control group (0 μg / mL) was set up, replacing the dacarbazin solution with an equal volume of sterile BHI broth. After 24 h of continuous culture, absorbance (OD) was measured at 600 nm every 1 h starting from the inception point. 600nm Finally, the growth time was used as the x-axis, and the OD (October Result) was calculated. 600nm Plot the 24-hour growth curve on the ordinate.

[0047] II. Experimental Results like Figure 2 As shown, dacarbazin exhibits antibacterial activity against Streptococcus mutans in vitro. At concentrations of 16 μg / mL and 64 μg / mL, dacarbazin nearly completely inhibited bacterial growth. The growth curves of each concentration treatment group in the figure are all separated from the solvent-based growth control group, indicating that dacarbazin can inhibit the normal growth process of Streptococcus mutans to varying degrees at different drug concentrations.

[0048] Example 3: Effects of dacarbazin on the amount and structural integrity of Streptococcus mutans biofilm formation. I. Biofilm biomass 1. Experimental Methods The Streptococcus mutans bacterial suspension was adjusted to 2×10⁻⁶ using BHIS medium. 6 After inoculating with CFU / mL, the cells were seeded into 96-well plates, and then an equal volume of dacarbazin solution was added to achieve final concentrations of 0 (control group), 4, 16, and 64 μg / mL, with three replicates per group. After 24 h of culture, the supernatant was discarded, and the cells were gently washed three times with PBS. The cells were then fixed with 4% paraformaldehyde solution (w / v) for 15 min, followed by staining with 0.1% crystal violet solution (w / v) for 5 min. After washing, anhydrous ethanol was added, and the crystal violet bound to the biofilm was dissolved at room temperature. The absorbance (OD) at 575 nm was measured using a microplate reader. 575 nm ).

[0049] 2. Experimental Results Crystal violet staining results showed that the crystal violet staining intensity was significantly reduced in the dacarbazine-treated experimental group, indicating a decrease in biofilm biomass. Figure 3At concentrations of 4, 16, and 64 μg / mL, dacarbazin inhibited the formation of Streptococcus mutans biofilm. The 4 μg / mL treatment group showed the highest OD... 575 nm The decrease was relatively small, but as the concentration increased to 16 and 64 μg / mL, OD... 575 nm The staining intensity decreased further, indicating a stronger inhibitory effect on biofilm formation. Figure 4 The biofilm formation in all three groups decreased significantly (P<0.05).

[0050] II. Bacterial aggregation and distribution within biofilms 1. Experimental Methods The Streptococcus mutans bacterial suspension was adjusted to 2×10⁻⁶ using BHIS medium. 6 After inoculating CFU / mL into 96-well plates, equal volumes of dacarbazin solution were added to achieve final concentrations of 0 (control group), 4, 16, and 64 μg / mL, with three replicates per group. The biofilm model was established after 24 h of incubation. The bacterial culture was discarded, and the plates were washed with PBS. Live / dead bacteria were stained using SYTO9 and PI fluorescent probes, and bacterial aggregation and fluorescence distribution within the biofilm were observed using an inverted fluorescence microscope.

[0051] 2. Experimental Results Live / dead staining results showed that the control group had a more continuous biofilm coverage and higher bacterial aggregation density; after dacarbazine treatment, the fluorescence distribution tended to be more dispersed, and the continuity of the biofilm decreased, with a more significant reduction in bacterial aggregation in the higher concentration groups. Both 16 and 64 μg / mL were higher than the MIC, and the decrease in biofilm formation may be jointly related to the inhibition of planktonic growth. Figure 5 ).

[0052] III. Integrity of Biofilm Structure 1. Experimental Methods Streptococcus mutans and dacarbazin were co-incubated on sterile slides. The final concentrations of dacarbazin in the incubation system were 0 (control group), 4, 16, and 64 μg / mL, respectively, and the slides were placed in 24-well plates. After 24 h of incubation, the slides were gently washed with PBS and fixed overnight at 4 °C with 2.5% glutaraldehyde solution (v / v). Subsequently, the slides were dehydrated with graded ethanol, critical point dried, and sputter-coated with gold. The ultrastructure of the biofilm was observed using scanning electron microscopy at accelerating voltages of 5 kV and magnifications of 5000× and 20000×.

[0053] 2. Experimental Results The ultrastructure of biofilms treated with different concentrations of dacarbazine was observed using scanning electron microscopy. In the control group, the biofilm exhibited a continuous and dense three-dimensional structure, with numerous bacteria tightly bound together and encapsulated by the extracellular matrix. When the dacarbazine concentration increased to 4 μg / mL, the biofilm structure gradually loosened and lacked integrity. At a concentration of 16 μg / mL, the biofilm further disintegrated, forming isolated colonies. At 64 μg / mL, the biofilm was almost completely removed, with only scattered free bacteria remaining. Figure 6 ).

[0054] The above-mentioned morphological changes are basically consistent with the results of crystal violet staining and live / dead staining, which together indicate that dacarbazin can disrupt the structural integrity of Streptococcus mutans biofilm in a concentration-dependent manner.

[0055] Example 4: Effects of dacarbazin on the metabolic activity and acid-producing capacity of Streptococcus mutans I. Metabolic Activity 1. Experimental Methods Streptococcus mutans was adjusted to 2 × 10⁻⁶ in BHIS medium. 6 After inoculating with CFU / mL, the cells were seeded into 96-well plates and cultured for 24 h to form a mature biofilm. After washing, dacarbazine solution was added and the cells were treated for another 24 h. The final concentrations of dacarbazine in the system were 0 (control group), 4, 16, and 64 μg / mL, respectively. Subsequently, 0.5 mg / mL MTT solution was added and incubated in the dark for 3 h. After discarding the supernatant, DMSO was added and shaken to dissolve the blue-purple crystalline formazan. The absorbance (OD) was measured at 490 nm. 490 nm ).

[0056] 2. Experimental Results The results showed that, compared with the control group, the amount of blue-purple crystalline formazan formed was significantly reduced after treatment with dacarbazine. Figure 7 Quantitative analysis showed that the metabolic activity of biofilms in each experimental group was significantly inhibited (P<0.05). Figure 8 The results indicate that dacarbazin can effectively interfere with the metabolic function of mature biofilms, and the metabolic inhibition effect is more significant with increasing compound concentration, showing a dose-dependent effect.

[0057] II. The ability of biofilms to produce lactic acid 1. Experimental Methods Streptococcus mutans was co-cultured with dacarbazin for 24 h, with final concentrations of dacarbazin of 0 (control group), 4, 16, and 64 μg / mL. After co-culture, the culture supernatant was discarded, and the culture was gently washed twice with PBS. 1 mL of buffered peptone solution was added, and the mixture was incubated anaerobicly at 37°C for another 2 h. The supernatant was collected, and the lactate content was determined according to the lactate assay kit instructions.

[0058] 2. Experimental Results like Figure 9 As shown, the control group had extremely high lactic acid production, while as the dacarbazine concentration increased to 64 μg / mL, the lactic acid concentration decreased in a dose-dependent manner. The differences between the experimental groups and the control group were statistically significant (P<0.05), demonstrating that dacarbazine can effectively inhibit the production of lactic acid by biofilms.

[0059] III. Acidity level of the culture environment 1. Experimental Methods The initial pH of the culture system containing dacarbazin was uniformly adjusted to 7.2. Subsequently, *Streptococcus mutans* was inoculated and cultured anaerobically at 37°C for 24 h. The final concentrations of dacarbazin in the culture system were 0 (control group), 4, 16, and 64 μg / mL, respectively. During the culture process, the pH value of each group was measured every 4 h using a precision pH meter. Each group was divided into three replicates to observe the degree of acidification of the culture environment caused by *Streptococcus mutans* metabolism under different treatment conditions.

[0060] 2. Experimental Results The results showed that dacarbazin significantly and dose-dependently inhibited the pH decrease caused by Streptococcus mutans metabolism within a certain concentration range. Figure 10 Within 24 hours, the pH value of the control group dropped sharply, falling below the cariogenic threshold of 5.5, while treatment with 64 μg / mL dacarbazine stabilized the pH value of the culture environment above a safe level, effectively inhibiting the formation of an acidic environment. The differences between the experimental groups and the control group were statistically significant (P<0.05).

[0061] Example 5: Effects of dacarbazin on the expression of genes related to biofilm formation in Streptococcus mutans I. Experimental Methods Streptococcus mutans was inoculated into BHIS medium and treated with dacarbazin solution for 24 h, with a final concentration of 64 μg / mL. An equal volume of solvent was added to the control group. After treatment, the supernatant was discarded, and TRIzol lysis buffer was immediately added to collect the biofilm. Total RNA was extracted after lysis with 100 μL of 30 mg / mL lysozyme for 30 min. RNA purity and concentration were determined spectrophotometrically, and qualified samples were used for reverse transcription to obtain cDNA. qRT-PCR was performed in 96-well plates using the SYBR Green system, with 16S rRNA as an internal reference gene. Target genes included... gtfB , gtfC , gtfD , srtA and gbpC The primer sequences are shown in Table 1, and the relative expression levels are calculated using a 23 ratio. - Ct Method calculation.

[0062] Amplification reaction program: 95℃ pre-denaturation for 30 s; 95℃ for 5 s, 60℃ for 30 s, for a total of 40 cycles; melting curves are used to determine amplification specificity.

[0063] Table 1 qRT-PCR primer sequences

[0064] II. Experimental Results Biofilm formation involves multiple related genes. qRT-PCR results showed that the expression of several key virulence genes in Streptococcus mutans was inhibited after treatment with dacarbazine. Figure 11 This includes not only gtfB, gtfC, and gtfD, which directly catalyze the synthesis of extracellular glucans, but also srtA, which is related to bacterial adhesion, and gbpC, which is related to stress response. The relative expression levels of these related genes were significantly decreased (P<0.05), which together led to impaired biofilm formation ability of Streptococcus mutans.

[0065] Example 6: Inhibitory effect of dacarbazine on dental caries in rats I. Experimental Methods Fifteen 17-day-old female SPF-grade SD rats were selected. From 17 to 20 days of age, the SD rats were administered 1 mg / mL ampicillin in their drinking water in conjunction with a complete diet to reduce oral endogenous flora. From 21 days of age, the SD rats were switched to Diet 2000 cariogenic diet and 5% sucrose water (w / v).

[0066] During the SD rat period of 21–28 days of age, the overnight activated Streptococcus mutans bacterial suspension was adjusted to 1×10⁻⁶. 8 CFU / mL, 0.2 mL of bacterial solution was applied to the oral cavity of each rat using a cotton swab, twice daily (morning and evening) for 7 consecutive days. After each application, the rats were deprived of food and water for 1 hour. Following infection, tooth samples from SD rats were locally treated and divided into a model group, a 1000 ppm NaF positive control group, and a 512 μg / mL dacarbazine treatment group, with 5 rats in each group. Model group: No drug intervention; NaF positive control group: Topical application of 1000 ppm NaF 0.2 mL / animal, twice daily at 12-hour intervals, for 4 consecutive weeks; Dacarbazine treatment group: 0.2 mL of 512 μg / mL dacarbazine solution was applied topically to each animal twice daily at 12-hour intervals for 4 consecutive weeks.

[0067] After the experiment, the upper and lower jaws were separated, fixed with 4% paraformaldehyde (w / v), and the mineralization structure and caries of the molars were observed using Micro-CT.

[0068] After cleaning, the jawbone specimens were immersed in a 0.4% ammonium purpurate solution (w / v) for 16 hours in the dark. The staining solution contained 4% paraformaldehyde (w / v). After staining, the specimens were rinsed with running water, dried at low temperature, and then hemisectioned along the midsagittal plane of the jawbone to expose the occlusal and proximal carious areas of each tooth. The red-stained areas were observed under a microscope, and the degree of caries was assessed according to the Larson Keyes scoring system (DOI: 10.1177 / 00220345580370060901). The scoring system included enamel lesions (E), minor dentin lesions (Ds), moderate dentin lesions (Dm), and extensive dentin lesions (Dx).

[0069] After euthanizing rats, buccal mucosa, gingiva, tongue, liver, spleen, and kidney tissues were isolated and washed with pre-cooled PBS, then fixed in 4% paraformaldehyde solution (w / v) for 24 h. The tissues were rinsed with running water, dehydrated with graded ethanol, cleared with xylene, and embedded in paraffin to prepare 4–5 μm serial sections. After dewaxing and rehydration, the sections were stained with H&E and observed under an upright optical microscope to examine the tissue structure and pathological changes.

[0070] II. Experimental Results The location of the caries is shown by the red arrow. Figure 12 When observed in the sagittal plane, the external contours and crown morphology of the teeth in each group were generally similar. The lesions in the model group were extensive, with dentin involvement in some areas. After treatment with dacarbazine and NaF, the surface structure of the enamel in the sagittal plane remained relatively intact, and no obvious deep lesions penetrating the entire enamel layer were observed.

[0071] Upon coronal observation, the differences in tooth mineralization structure among the groups were minimal. A defect was observed in the model group, indicating demineralization damage to the hard tissues of the tooth after modeling. Compared to the model group, the lesion area in the dacarbazine-treated group was smaller, and the depth of the radiolucent zone was shallower. The NaF-positive control group showed intact and dense tooth structure, with mineralization closest to normal teeth. The results indicate that 512 μg / mL dacarbazine can also reduce enamel damage to molars caused by caries in animals, demonstrating good anti-caries efficacy.

[0072] The lesion area after ammonium purpurate staining is shown by the black arrow. Figure 13 The model group rats showed obvious caries lesions in the molar fissures; the caries lesions in the molar fissures of the dacarbazine treatment group and the NaF positive control group were smaller than those in the model group, showing a more obvious anti-caries effect and effectively inhibiting the progression of caries in rat tooth tissue.

[0073] On the four scoring indicators of enamel caries (E), mild dentin caries (Ds), moderate dentin caries (Dm), and severe dentin caries (Dx), the scores of the dacarbazine treatment group and the NaF positive control group were lower than those of the model group. Figure 14 Among them, the reduction in the scores of Ds, Dm, and Dx was particularly significant. The scores of the dacarbazine treatment group were close to those of the NaF positive control group, preventing the caries from progressing to deeper layers.

[0074] H&E staining showed that the buccal mucosa, gingiva, tongue, liver, spleen, and kidney tissues of the model group, dacarbazine treatment group, and NaF positive control group were generally intact, with no obvious abnormalities in cell morphology. No obvious inflammatory cell infiltration, tissue edema, cell degeneration and necrosis, or fibrous tissue hyperplasia were observed.

[0075] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. Application of dacarbazin in the preparation of inhibitors of Streptococcus mutans.

2. Application of dacarbazin in the preparation of inhibitors for biofilm formation of Streptococcus mutans.

3. The use of dacarbazin in the preparation of drugs for the prevention and / or treatment of Streptococcus mutans infection.

4. Application of dacarbazin in the preparation of drugs that inhibit the expression of gtfB, gtfC, gtfD, gbpC and / or srtA genes in Streptococcus mutans.

5. Use of dacarbazin in the preparation of drugs for the prevention, improvement and / or treatment of diseases caused by Streptococcus mutans.

6. Use of dacarbazine in the preparation of drugs for the prevention, improvement and / or treatment of dental caries.

7. The application according to claim 6, characterized in that, The drug inhibits the growth of Streptococcus mutans and / or inhibits the formation of Streptococcus mutans biofilm.

8. The application according to any one of claims 1 to 6, characterized in that, The effective concentration of dacarbazine is 3–70 μg / mL.

9. The application according to claim 8, characterized in that, The effective concentration of dacarbazine is 4–64 μg / mL.

10. The application according to any one of claims 3 to 6, characterized in that, The drug also includes pharmaceutically acceptable excipients.

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