CN116509851B - Use of Ha Erfen in preparation of herpes simplex virus inhibitor - Google Patents
Use of Ha Erfen in preparation of herpes simplex virus inhibitor Download PDFInfo
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- CN116509851B CN116509851B CN202310392717.4A CN202310392717A CN116509851B CN 116509851 B CN116509851 B CN 116509851B CN 202310392717 A CN202310392717 A CN 202310392717A CN 116509851 B CN116509851 B CN 116509851B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
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- A—HUMAN NECESSITIES
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
- A61P31/22—Antivirals for DNA viruses for herpes viruses
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention provides an application of halphenol in preparing a herpes simplex virus inhibitor, and experimental results prove that Ha Erfen has an inhibiting effect on herpes simplex virus type I (HSV-1) and also has a strong inhibiting effect on acyclovir drug-resistant strains (HSV-1/153), can effectively inhibit virus replication and improve survival rate of infected cells, and in vitro cells and animal experiments show that the halphenol has lower toxicity on corneal epithelial cells and mouse corneas and has a good treatment effect on herpes simplex virus keratitis.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to application of halphenol in preparation of a herpes simplex virus inhibitor.
Background
Herpes simplex virus keratitis (herpes simplex keratitis, HSK) caused by invasion of cornea tissue by herpes simplex virus I (herpes simplex virus-1, HSV-1) has the characteristics of high incidence, high treatment difficulty, high recurrence rate and the like, and is one of the main blindness-causing eye diseases at present. The primary HSV infection is mostly asymptomatic infection, can be hidden in ganglions after the primary infection, and can be activated again under certain conditions to cause recurrent infection. Recurrent infections can cause more severe corneal scarring and neovascularization, ultimately leading to corneal haze and even blindness. HSK is thus one of the major public health and social problems at present.
Acyclovir is currently the first drug to treat HSV infection, but acyclovir has the following problems during HSK treatment: (1) Frequent topical administration and blurred vision after acyclovir gel use make it difficult for patients to follow the order; (2) Prolonged use of drugs has led to the emergence of acyclovir resistant strains; (3) there is currently no effective vaccine against HSV infection. Therefore, there is an urgent need to find a new compound having different pharmacological actions to solve the above problems.
Disclosure of Invention
The invention aims to provide an application of halphenol in preparation of a herpes simplex virus inhibitor based on the prior art.
A second object of the present invention is to provide the use of Ha Erfen as described above for the preparation of a medicament as a herpes simplex virus inhibitor.
A third object of the present invention is to provide an application of Ha Erfen in the preparation of a medicament for treating a disease caused by herpes simplex virus type I (HSV-1), for example, herpes simplex keratitis.
The fourth object of the invention is to provide the application of Ha Erfen in preparing the medicament for treating diseases caused by acyclovir-resistant strain herpes simplex virus type I/153 (HSV-1/153).
A fifth object of the present invention is to provide a pharmaceutical composition comprising haversol as an active ingredient or a major active ingredient, together with a pharmaceutically acceptable carrier.
The technical scheme of the invention is as follows:
Use of Ha Erfen in the preparation of a herpes simplex virus inhibitor, wherein Ha Erfen has the structural formula:
in a preferred embodiment, ha Erfen is used in the preparation of a medicament as a herpes simplex virus inhibitor.
When Ha Erfen is applied to the preparation of the medicament serving as the herpes simplex virus inhibitor, the medicament can be prepared into a solid preparation or a liquid preparation. Furthermore, on the basis of the technical scheme provided by the invention, the medicine can be prepared into injection, oral liquid, granules, powder, tablets, capsules or external patches.
Haer phenol is a beta-carboline alkaloid, and the molecular formula is as follows: c 12H10N2O, molecular weight: 198.22100 has various pharmacological effects, and no research report on inhibiting HSV-1 or HSV-1/153 by Ha Erfen exists at present, and no report on application of Ha Erfen in preparing medicaments for treating herpes simplex keratitis exists.
In the invention, ha Erfen not only can inhibit the expression of HSV-1 envelope protein gD-1, but also can inhibit the expression of acyclovir resistant strain HSV-1/153 envelope protein gD-1, thereby inhibiting the replication of HSV-1 and HSV-1/153, and having obvious antiviral effect.
In a preferred embodiment, ha Erfen is used in the manufacture of a medicament for the treatment of a disease caused by herpes simplex virus type I. For example, the disease caused by herpes simplex virus type I is herpes simplex keratitis.
In a preferred embodiment, ha Erfen is used in the manufacture of a medicament for the treatment of a disease caused by the acyclovir-resistant strain herpes simplex virus type I/153.
In the invention, experimental results prove that Ha Erfen not only has an inhibiting effect on herpes simplex virus type I (HSV-1), but also has a strong inhibiting effect on acyclovir drug-resistant strains (HSV-1/153), can effectively inhibit viral replication and improve the survival rate of infected cells, and in vitro cells and animal experiments show that the halphenol has lower toxicity on corneal epithelial cells and mouse corneas and has a good treatment effect on herpes simplex keratitis.
The invention also provides a pharmaceutical composition which takes the halphenol as an active ingredient or a main active ingredient and is assisted with a pharmaceutically acceptable carrier. The pharmaceutical composition can be solid preparation or liquid preparation, in particular injection, oral liquid, granule, powder, tablet, capsule or external patch. Pharmaceutically acceptable auxiliary materials play a key role in the whole preparation technology development process, and the auxiliary materials account for most of the preparation, so that the properties of the auxiliary materials determine the properties of the preparation to a great extent. The good auxiliary materials can enhance the stability of the main medicine and prolong the effective period of the medicine; the release speed of the main medicine in vivo and in vitro can be regulated and controlled; can change the absorption of the medicine in the body and increase the bioavailability, and the specific auxiliary material type is not limited.
By adopting the technical scheme of the invention, the advantages are as follows:
the invention provides application of halphenol in preparation of a herpes simplex virus inhibitor, and experimental results prove that Ha Erfen has an inhibiting effect on herpes simplex virus type I (HSV-1) and also has a strong inhibiting effect on acyclovir drug-resistant strains (HSV-1/153), can effectively inhibit virus replication and improve survival rate of infected cells, and in vitro cells and animal experiments show that the halphenol has lower toxicity on corneal epithelial cells and mouse cornea and has a good treatment effect on herpes simplex virus keratitis.
Drawings
FIG. 1 is a graph showing the results of the In-cell Western method test Ha Erfen (Harmol) of example 1 of the present invention for inhibiting HSV-1 and HSV-1/153 infection on corneal epithelial cells (HCEC cells), wherein (A) In FIG. 1 is an immunofluorescence photograph of HSV-1 envelope proteins gD-1 and DRAQ5 (internal control normalization) after treatment of HCEC cells infected with HSV-1 with different concentrations of haversol; in the figure 1 (B), immunofluorescence reading values of gD-1 and DRAQ5 are read through Oddssey software, the gD-1/DRAQ5 ratio is obtained through calculation, gD-1/DRAQ5 of a virus control group treated by 0.02% DMSO is self-defined to be 1.0, and the numerical values of gD-1/DRAQ5 are calculated respectively by analogy with virus groups treated by haversol with different concentrations; FIG. 1 (C) is an immunofluorescence photograph of HSV-1/153 envelope proteins gD-1 and DRAQ5 (reference standardized) after treatment of HCEC cells infected with HSV-1/153 with different concentrations of haversol; in FIG. 1, (D) is immunofluorescence reading of gD-1 and DRAQ5 by Oddssey software, the gD-1/DRAQ5 ratio is obtained, gD-1 of a virus control group treated by 0.02% DMSO is defined as 1.0, and the values of gD-1/DRAQ5 are calculated by analogy with virus groups treated by haversol with different concentrations.
FIG. 2 is a graph showing the results of the In-cell Western assay of example 2 of the present invention for inhibition of HSV-1 and HSV-1/153 infection by HSV-cell-Western assay Ha Erfen (Harmol) on corneal epithelial cells (HCEC cells), wherein (A) In FIG. 2 is the half-effective inhibitory concentration (EC 50) of Ha Erfen (Harmol) for inhibition of HSV-1 infection by HCEC cells; FIG. 2 (B) is a half-effective inhibitory concentration (EC 50) of Ha Erfen (Harmol) for inhibiting HSV-1/153 infection of HCEC cells.
FIG. 3 is a graph showing the results of inhibiting infection of corneal epithelial cells (HCEC cells) with HSV-1 and HSV-1/153 using Vero cell assay Ha Erfen in example 3 of the present invention, wherein (A) in FIG. 3 is virus titer (TCID 50) after re-infection of Vero cells with cell supernatant by the Reed-Muench method after treatment of HCEC infected with HSV-1 with different concentrations of Harnol (Harmol); FIG. 3 (B) shows virus titers (TCID 50) of the cell supernatants after reinfection with Vero cells by the Reed-Muench method after treatment with varying concentrations of Harhenol (Harmol) to infect HCEC with HSV-1/152.
FIG. 4 shows cytotoxicity of haversol (Harmol) on HCEC cells at various concentrations in example 4 of the present invention.
FIG. 5 shows the efficacy of Ha Erfen (Harmol) of example 5 of the present invention in treating herpes simplex keratitis caused by HSV-1 infected mouse cornea; wherein, (a) in fig. 5 is a corneal slit lamp picture; fig. 5 (B) is a score chart of the slit lamp evaluation blepharitis; fig. 5 (C) corneal fluorescein sodium stained slit lamp picture; fig. 5 (D) shows the relative defect area of the corneal epithelium; in FIG. 5 (E) is HSV-1 virus titer (TCID 50) in mouse tears.
Detailed Description
The invention will be better understood from the following examples. However, it will be readily appreciated by those skilled in the art that the description of the embodiments is provided for illustration only and should not limit the invention as described in detail in the claims.
The material information used in the embodiments of the present invention is as follows:
herpes simplex virus type I (HSV-1F strain) and herpes simplex virus type I (HSV-1/153 strain) were purchased from American Type Culture Collection (ATCC).
African green monkey kidney cells (Vero cells) and corneal epithelial cells (HCEC) were purchased from the national academy of sciences cell bank.
HSV-1gD antibody, available from St. Kruese Biotechnology Co., ltd., goat anti-mouse secondary antibody (IRDye 800CW Goat-anti-mouse 800) was purchased from LI-COR (Lincoln, NE, USA) Inc. in the United states.
Ha Erfen (Harmol) was purchased from small molecule compounds and natural product libraries (Shanghai, china).
Example 1
Antiviral effect of Ha Erfen on inhibiting infection of HCEC cells with HSV-1 or HSV-1/153 by In-cell-Western method
HCEC cells were inoculated in 96-well plates, each well containing 100 μl of fresh complete medium, and when cell density reached above 90%, halphenol (Ha Erfen in DMSO) at concentrations of 3.125, 6.25, 12.5, 25, 50 μm was added, respectively, and incubated in a cell incubator at 37 ℃ with 5% co 2 for 30min; HCEC cells were then infected with HSV-1 and HSV-1/153 (infectious virus titer moi=1), respectively; after 24h, fixing with 4% paraformaldehyde for 30min, and adding 200 μl of 0.1% Triton X-100 buffer into each well for penetration treatment for 20min; sealing for 90min by using 4% skimmed milk powder after the penetration is finished, and adding primary antibody (1:200) of envelope protein gD into each hole for incubation for 2h; washing the plate with PBST buffer solution for 5 times, 5min each time, adding secondary antibody (1:2500) and DRAQ5 into each hole, and incubating for 1h; the plates were washed 5 times with PBST buffer for 5min each and finally tested with Oddssey INFRARED IMAGE SYSTEM.
The results are shown in FIG. 1: ① gD-1 protein expression on the longitudinal axes of HSV-1 and HSV-1/153 groups was 0.17 and 0.08, respectively, at a haversol concentration of 3.125. Mu.M on the transverse axis; ② gD-1 protein expression on the longitudinal axes of HSV-1 and HSV-1/153 groups was 0.13 and 0.06, respectively, at a haversol concentration of 6.25. Mu.M on the transverse axis; ③ At a haer phenol concentration of 12.50. Mu.M, the gD-1 protein expression on the vertical axes of HSV-1 and HSV-1/153 groups was 0.09 and 0.04, respectively; ④ gD-1 protein expression on the longitudinal axes of HSV-1 and HSV-1/153 groups was 0.06 and 0.03, respectively, at a haversol concentration of 25. Mu.M on the transverse axis; ⑤ On the transverse axis, at a haversol concentration of 50. Mu.M, the gD-1 protein expression on the longitudinal axes of HSV-1 and HSV-1/153 groups was 0.04 and 0.01, respectively. The expression degree of HSV envelope protein gD-1 is in direct proportion to the replication level of progeny virus, which shows that the haversol can inhibit the replication of HSV-1 in HCEC cells, can inhibit the replication of acyclovir resistant strain HSV-1/153, and is in a dose-dependent curve relationship. The HCEC cells are human cornea epithelial cells and can be used for simulating a disease model of herpes simplex keratitis caused by the infection of eyes by HSV-1 and HSV-1/153 viruses in a cell experiment. Harpolol inhibits replication of HSV-1 and HSV-1/153 in corneal epithelial cells, demonstrating Ha Erfen can be used for treatment of herpes simplex keratitis caused by HSV-1 and HSV-1/153 infection.
Example 2
Detection of Ha Erfen and acyclovir inhibition of infection of HCEC cells by HSV-1 and HSV-1/153 by In-cell-Western method
HCEC cells were inoculated in 96-well plates, each well containing 100 μl of fresh complete medium, and when cell density reached above 90%, halphenol (Ha Erfen in DMSO) at concentrations of 3.125, 6.25, 12.5, 25, 50 μΜ and acyclovir 10 μΜ were added and incubated in a cell incubator at 37 ℃ with 5% co 2 for 30min, respectively; vero cells were then infected with HSV-1 and HSV-1/153 (infectious virus titer moi=1), respectively; after 24h, fixing with 4% paraformaldehyde for 30min, and adding 200 μl of 0.1% Triton X-100 buffer into each well for penetration treatment for 20min; sealing for 90min by using 4% skimmed milk powder after the penetration is finished, and adding primary antibody (1:200) of envelope protein gD into each hole for incubation for 2h; washing the plate with PBST buffer solution for 5 times, 5min each time, adding secondary antibody (1:2500) and DRAQ5 into each hole, and incubating for 1h; the PBST buffer is washed for 5 times, each time for 5min, and finally Oddssey INFRARED IMAGE SYSTEM is used for detection, and the inhibition rate of viruses is calculated as follows: [ 1-expression of gD protein in drug-treated group/expression of gD protein in control group ]. Times.100%.
As shown in FIG. 2, ① on the horizontal axis, at a halphenol concentration of 3.125. Mu.M, the inhibition rates of HSV-1 and HSV-1/153 on the vertical axis were 22.24% and 36.45%, respectively; ② On the horizontal axis, at a halphenol concentration of 6.25. Mu.M, the HSV-1 and HSV-1/153 inhibition rates on the vertical axis were 39.85% and 45.74%, respectively; ③ On the horizontal axis, at a halphenol concentration of 12.50. Mu.M, the inhibition rates of HSV-1 and HSV-1/153 on the vertical axis were 57.75% and 70.82%, respectively; ④ On the horizontal axis, HSV-1 and HSV-1/153 inhibition rates on the vertical axis were 72.85% and 77.65%, respectively, at a halphenol concentration of 25. Mu.M; ⑤ On the horizontal axis, HSV-1 and HSV-1/153 inhibition rates on the vertical axis were 85.35% and 82.55% respectively at a halphenol concentration of 50. Mu.M; ⑥ On the horizontal axis, HSV-1 and HSV-1/153 inhibition rates on the vertical axis were 78.27% and 9.65%, respectively, at acyclovir concentrations of 10. Mu.M.
Conclusion: the half-effective inhibition concentrations EC 50. Ha Erfen calculated on the horizontal axis according to Graphpad statistical software inhibited HSV-1 and HSV-1/153 replication at EC 50 of 8.25 and 5.84. Mu.M, respectively. By comparing the sizes of EC 50, the replication effect of the haversol on inhibiting HSV-1/153 is better than that of the haversol on inhibiting HSV-1. The results show that the haversol has a strong inhibition effect on acyclovir resistant strains HSV-1/153, and can be used for treating diseases caused by acyclovir resistant strain infection.
Example 3
Detection Ha Erfen of infection of corneal epithelial cells (HCEC cells) with HSV-1 and HSV-1/153 using Vero cells
HCEC cells were inoculated in 96-well cell culture plates, after cell confluence, ha Erfen (Ha Erfen in DMSO) and 10 μm acyclovir were added at concentrations of 0, 6.25, 12.5, 25 μm, respectively, and after incubation for 30min at room temperature, HCEC cells were infected with HSV-1 and HSV-1/153 (moi=1); after 24h, the medium was discarded, 200. Mu.L of fresh medium was added, and then repeated freeze thawing was performed 3 times at-20℃and 37℃to release the virus particles. The virus particles contained in the fresh culture medium are infected with Vero cells, the Vero cells are inoculated in a 96-well plate in advance for 24 hours, 50 mu L of the culture medium containing the virus particles is added into the wells after the virus particles are converged, and after 72 hours, the numbers of HSV-1 and HSV-1/153 virus plaques are recorded and counted respectively under an optical microscope.
As a result, FIG. 3 shows that ① on the horizontal axis and 5.8 and 5.9 on the vertical axis at a haversol concentration of 0. Mu.M; ② On the horizontal axis, at a haversol concentration of 6.25. Mu.M, the numbers of HSV-1 and HSV-1/153 plaques on the vertical axis are 5.1 and 5.0, respectively; ③ On the horizontal axis, HSV-1 and HSV-1/153 plaque numbers on the vertical axis are 4.6 and 4.0, respectively, at a haversol concentration of 12.5. Mu.M; ④ On the horizontal axis, HSV-1 and HSV-1/153 plaque numbers on the vertical axis are 4.5 and 3.1, respectively, at a halphenol concentration of 25. Mu.M; ⑤ On the horizontal axis, HSV-1 and HSV-1/153 plaque numbers on the vertical axis were 4.2 and 5.8, respectively, at acyclovir concentration of 10. Mu.M.
Conclusion: the numerical values of the HSV-1 and HSV-1/153 plaque numbers are in direct proportion to the replication level of the progeny virus, which shows that the halphenol can inhibit the infection of HCEC by the HSV-1 progeny virus, the inhibiting effect of the haliol on the HSV-1 is better than that of 10 mu M acyclovir at Ha Erfen concentration of more than 25 mu M, and the Ha Erfen has obvious inhibiting effect on acyclovir resistant strains.
Example 4 Ha Erfen cytotoxicity test against HCEC
HCEC cells were seeded in 96-well cell culture plates and, after cell confluence, gradient concentrations (0, 6.25, 12.5, 25, 50, 100, 150, 200, 250 μm) of Ha Erfen were added, respectively; after 72h incubation in a cell incubator with 5% CO 2 at 37℃10 μl CCK8 was added per well and incubated in a cell incubator with 5% CO 2 at 37℃for 3h; finally, the absorbance size (OD) of each well at 450nm was measured using a TECAN instrument. The cell survival ratio is the ratio of the absorbance of the cells of the wells of the Galhalphenol to the absorbance of the cells of the wells without the addition of Ha Erfen.
The results are shown in FIG. 4, in which HCEC cells were treated with different concentrations of halphenol on the horizontal axis; on the vertical axis, the cell viability of HCEC cells after treatment with haversol at different concentrations was divided. In vitro cytotoxicity experiments show that half cytotoxicity concentration (half cytotoxicity concentration, CC 50) of haer phenol on HCEC cells is 260 mu M, which is obviously lower than the use concentration in each experiment, so that the inhibition effect of Ha Erfen on HSV-1 and HSV-1/153 is not caused by the cytotoxicity effect of the haer phenol on the cells.
Example 5 animal experiments
18 BALB/c males at 6 weeks of age were selected and randomly divided into 3 groups: 6 were used as control groups (MOCK) and 12 were used as infected groups. Anesthesia was given by intraperitoneal injection with 1% sodium pentobarbital (80 mg/kg), respectively; selecting the right eye of a mouse as a study object, and making a "#" scratch on the surface of the cornea by using a 27G injection needle to penetrate the corneal epithelium; the corneal surface was instilled with 5. Mu.L of DMEM with or without HSV-1 (1X 10 6 PFU/ml). Of these, 6 mice were inoculated with DMEM containing no HSV-1 as a control group, and 12 mice were inoculated with DMEM containing HSV-1 as an infected group. The 12 infected mice were randomly divided into two groups; starting 24 hours after infection, the medicine is administrated three times a day for continuous 5 days, and 5 mu L of the medicine is dripped on the cornea surface each time; wherein the control group was treated with PBS, the infected group was treated with PBS, and the other group was treated with haversol. Slit lamps were used to observe the relative area of corneal epithelial defects, blepharitis extent, and HSV-1 virus titer in mouse tears at day 3 and day 5 after treatment, respectively, using the TCID 50 method. The scoring criteria for blepharitis (blepharitis score) were: 0 point: edema of eyelid margin; 1, the method comprises the following steps: mild oedema of the eyelid margin; 2, severe eyelid edge edema with severe eyelid crusting; 3, the method comprises the following steps: eyelid half closure is accompanied by severe eyelid margin crusting; 4, the following steps: the eyelid is completely closed. Corneal epithelium versus defect area (RELATIVE DEFECT) ImageJ software measures the ratio of the area of corneal epithelial sodium fluorescein staining to the total corneal area x 100%. The major clinical symptoms of HSK include corneal edema, epithelial defects, blepharitis, where the more pronounced corneal edema indicates a heavier symptom, the larger the area of corneal epithelial defects indicates a heavier symptom, and the higher the blepharitis score value indicates a heavier symptom.
The results are shown in fig. 5, with ① haar phenol treatment group cornea clear without significant corneal edema; ② The severity of haversol-treated blepharitis was significantly lower than in the PBS-treated group; ③ The area of the corneal epithelial defect of the haversol treatment group is obviously smaller than that of the PBS treatment group; ④ The relative defect area of the cornea epithelium of the haer phenol group is obviously lower than that of the PBS treatment group; ⑤ HSV-1 virus titers in tears of the Harpoled treated mice were significantly lower than in the PBS treated mice.
Conclusion: the haversol can obviously inhibit the replication of HSV-1 in mouse HSK, and effectively lighten the damage of HSV-1 to mouse cornea, thereby obviously relieving the clinical symptoms of HSK.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments may be modified or some technical features may be replaced equivalently; such modifications and substitutions do not depart from the spirit of the invention.
Claims (3)
1. Use of Ha Erfen in the manufacture of a medicament for the treatment of a disease caused by the herpes simplex virus type I/153 of an acyclovir-resistant strain.
2. The use according to claim 1, wherein the medicament is formulated as a liquid or solid formulation.
3. The use according to claim 2, wherein the medicament is formulated as an injection, an oral liquid, a granule, a powder, a tablet, a capsule or a patch for external use.
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| 骆驼蓬生物总碱体外抗病毒作用的实验研究;杨红， 陶佩珍;江苏中医药;20060105;第27卷(第1期);第54-56页 * |
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