Method for treating a glycoprotein-related disease

Abstract

A method for treating a glycoprotein-related disease is disclosed, which comprises: administering a first effective amount of phenol red and a second effective amount of an organic arsenic compound to a subject in need thereof.

Claims

1. A method for treating a glycoprotein-related disease, comprising: administering a first effective amount of phenol red and a second effective amount of an organic arsenic compound to a subject in need thereof; wherein the glycoprotein-related disease includes one caused by human immunodeficiency virus.

2. The method of claim 1, wherein the organic arsenic compound is selected from the group consisting of monosodium methanearsonate, methylarsonic acid, sodium dimethylarsonate, disodium methylarsonate, cacodylic acid, and calcium acid methanearsonate.

3. The method of claim 1, wherein a pharmaceutical acceptable carrier is further administered to the subject in need thereof.

4. The method of claim 3, wherein the pharmaceutical acceptable carrier is selected from the group consisting of solvent, buffer, suspending agent, decomposer, disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, lubricant, absorption delaying agent, and liposome.

5. The method of claim 1, wherein the first effective amount is ranged from 0.1 mg to 5.0 mg per kilograms of the subject in need thereof.

6. The method of claim 5, wherein the first effective amount is ranged from 0.5 mg to 1.5 mg per kilograms of the subject in need thereof.

7. The method of claim 1, wherein the first effective amount is ranged from 0.1 mg/cc to 5.0 mg/cc.

8. The method of claim 1, wherein the first effective amount is ranged from 0.8 mg/cc to 3.0 mg/cc.

9. The method of claim 1, wherein the second effective amount is ranged from 0.1 mg to 10.0 mg per kilograms of the subject in need thereof.

10. The method of claim 9, wherein the second effective amount is ranged from 0.8 mg to 2.0 mg per kilograms of the subject in need thereof.

11. The method of claim 1, wherein the second effective amount is ranged from 0.1 mg/cc to 10.0 mg/cc.

12. The method of claim 11, wherein the second effective amount is ranged from 2.0 mg/cc to 6.0 mg/cc.

13. A method for preventing a disease caused by human immunodeficiency virus, comprising: providing an article in contact with a subject in need; wherein the article contains a pharmaceutical composition comprising phenol red and an organic arsenic compound; wherein the pharmaceutical composition prevents the disease caused by human immunodeficiency virus.

14. The method of claim 13, wherein the article comprises one selected from a group consisting of a condom, an ointment, a lotion and a medical tool.

15. The method of claim 14, wherein the medical tool comprises one selected from a group consisting of a syringe, a gauze, a medical tube and a surgical tool.

16. The method of claim 13, wherein the organic arsenic compound is selected from the group consisting of monosodium methanearsonate, methylarsonic acid, sodium dimethylarsoriate, disodium methylarsonate, cacodylic acid, and calcium acid methanearsonate.

17. The method of claim 13, wherein the pharmaceutical composition further comprises a pharmaceutical acceptable carrier.

18. The method of claim 17, wherein the pharmaceutical acceptable carrier is selected from the group consisting of solvent, buffer, suspending agent, decomposer, disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, lubricant, absorption delaying agent, and liposome.

19. The method of claim 13, wherein the subject is a mammal.

20. The method of claim 19, wherein the mammal is a human being.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a figure showing the leukocyte amount changes of the FIV infected cat during the treatment according to Embodiment 1 of the present disclosure;

(2) FIG. 2 is a figure showing MAGIC-5 Cell viability assay of LuMC5;

(3) FIG. 3 is a figure showing drug susceptibility assay of LuMC5 treated in HIV-1 CRF07_BC infection model.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

(4) The present invention has been described in an illustrative manner, and it is to be understood that the terminology used is intended to be in the nature of description rather than of limitation. Many modifications and variations of the present invention are possible in light of the above teachings. Therefore, it is to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described.

(5) In the following embodiments, a solution containing phenol red (2 mg/cc) and monosodium methanearsonate (4 mg/cc) is used, which is named as LuMC5, hereinafter. In addition, a solution containing monosodium methanearsonate (4 mg/cc) alone is also used, which is named as MASA, hereinafter. It should be noted that the following embodiments only provide one formulation of the pharmaceutical composition of the present disclosure. A person skilled in the art knows that the concentration of the active ingredients and the contents of the pharmaceutical composition can be adjusted within the scope of the appended claims.

Embodiment 1

(6) In the present embodiment, feline immunodeficiency virus (FIV), which is belonged to the same family (Retroviridae) and the same genus (Lentivirus) to HIV, was selected to perform the present clinical experiment. LuMC5 was administered to the infected cat via intravenous injection or intramuscular injection, 0.4 cc/kg, once or twice a day. The results are shown in the following Table 1 and FIG. 1.

(7) TABLE-US-00001 TABLE 1 Blood cells examination data of FIV infected cat during the treatment (Testing company: LEZEN Reference Lab) WBC NEU LYM MONO EOSI BASO Day 1 530 18.9 43.4 37.7 0 0 Day 3 12790 73 10 6.6 0.3 0.1 Day 9 31670 80 9 6 4.9 0.1 Day 34 9000 69 24 5 2 0.1 RBC Hgb HCT M.C.V M.C.H M.C.H.C PLT Day 1 7.84 12.1 44.5 56.8 15.4 27.2 56 Day 3 7.7 11.9 42.1 54.7 15.5 28.3 165 Day 9 6.82 10.6 34.8 51 15.5 30.5 132 Day 34 6.71 10.2 37.5 55.9 15.2 27.2 38 NEU: Neutrophils/ LYM: Lymphocytes/ MONO: Monocytes/ EOSI: Eosinophils/ BASO: Basophils/ Hg: Hemoglobin/ HCT: Hematocrit/

(8) From the results shown in FIG. 1 and Table 1, the immune cell amount was increased rapidly at Day 3. In addition, acquired immune deficiency syndromes caused by FIV, such as toxoplasmosis, pneumocystis pneumonia and tumors were not found. The FIV infected cat turned into a healthy carrier after treatment.

(9) In addition, LuMC5 can be administered to the FIV infected cat for 4 to 14 days. No virus is detected after 4 to 14 days. Hence, LuMC5 can be administered when the virus levels in the FIV infected cat is high to control the syndromes, and is not have to be administered for a long term. Therefore, the immune system of the FIV infected cat is not destroyed.

(10) Therefore, when LuMC5 is used for treating the FIV infected subject, the development of the acquired immune deficiency syndromes caused by FIV can be inhibited; and the complication such as toxoplasmosis and pneumocystis pneumonia can be inhibited. Hence, the purposes of treatment and prevention can be accomplished.

Embodiment 2

(11) In the present embodiment, MASA and LuMC5 were used to treat the virus infected vertebrate and mammalian. 7 virus species belonged to 5 families were selected to perform the present clinical experiment. Herein, the virus classification is on the basis of National Center for Biotechnology Information, U. S.

(12) Subject: Infected dogs and cates sent to Hei-Ming Veterinary Hospital, Taiwan

(13) 7 virus species for clinical experiments:

(14) Family: Retroviridae

(15) Genus: Gammaretrovirus species: Feline leukemia virus Genus: Lentivirus species: Feline immunodeficiency virus
Family: Parvoviridae species: Canine parvovirus species: Feline panleukopenia virus
Family: Paramyxoviridae Genus: Morbillivirus species: Canine distemper virus
Family: Coronaviridae Genus: Alphacoronavirus species:Feline coronavirus
Family: Herpesviridae Genus: Varicellovirus species: Felid herpesvirus 1
Treatment: Intravenous injection or intramuscular injection, 0.4 cc/kg, once or twice a day Testing company: Genomics BioSci & TechCo., Ltd. (Abbreviation: G) MountainVet Biotech Co., Ltd. (Abbreviation: M) VETE Co., Ltd. (Abbreviation: V) LEZEN Reference Lab (Abbreviation: L)
Testing instrument: Real-time quantitative PCR detecting system (sensitivity >99%) for G and M companies; and PCR or RT-PCR for V company
Testing method: The detected genes are the long terminal repeat (LTR) gene of Feline immunodeficiency virus, the virus protein 2 (VP2) gene of Canine parvovirus and the nucleocapsid gene (N gene) of Canine distemper virus. The virus amount is a curve caluated by the cycle threshold (C.sub.t) values of the obtained PCR products based on the cycle threshold (C.sub.t) values of the standard with predetermined different concentrations.

(16) The testing results of the aforementioned 7 viruses in the clinical experiments are listed in the following Tables 2-1 to 2-7.

(17) TABLE-US-00002 TABLE 2-1 Results of the clinical experiments on Feline leukemia virus Animal Total Detecting Results & ID Age Gender Medicine Days point (ng/uL) Specimen Company Cat 2 M MSMA 9 Day 0 + (*.sup.a) Blood V 130730 Years Day 9 (*.sup.b) Cat 7 F LuMC5 9 Day 0 222.8 10.sup.7 Blood M 160312 Years Day 11 98.75 10.sup.7 Cat 1 F LuMC5 7 Day 0 248.8 10.sup.7 Blood M 150903 Years Day 9 87.51 10.sup.7 Cat 2 F LuMC5 6 Day 0 261.4 10.sup.7 Blood M 160729 Months Day 4 74.6 10.sup.7 Day 6 53.6 10.sup.7

(18) TABLE-US-00003 TABLE 2-2 Results of the clinical experiments on feline immunodeficiency virus Animal Total Detecting Results & ID Age Gender Medicine Days point (ng/uL) Specimen Company Cat 2 M MSMA 9 Day 0 + (*.sup.a) Blood V 130730 Years Day 9 (*.sup.b) Cat 7 F MSMA 14 Day 0 + (*.sup.a) Blood V 130805 Months Day 14 (*.sup.b) Cat 7 F LuMC5 9 Day 0 45.75 10.sup.7 Blood M 160312 Years Day 11 Cat 2 F LuMC5 6 Day 0 34.8 10.sup.7 Blood M 160729 Months Day 4 Day 6

(19) TABLE-US-00004 TABLE 2-3 Results of the clinical experiments on canine parvovirus Animal Total Detecting Results & ID Age Gender Medicine Days point (ng/uL) Specimen Company Dog 6 F MSMA 4 Day 0 932918 10.sup.8 Feces (*c) G 140529 Months Day 4 28836 10.sup.8 Dog 3 F MSMA 10 Day 0 458306 10.sup.8 Blood M 150716 Months Day 10 2470 10.sup.8 Feces (*d) Dog 2 F MSMA 10 Day 0 40724.24 10.sup.7 Blood M 150708 Months Day 8 2696.9 10.sup.7 Day 15 185.6 10.sup.7 Dog 1 M MSMA 7 Day 0 46322 10.sup.8 Blood G 150125 year Day 3 345 10.sup.8 Day 5 75 10.sup.8 Day 6 40 10.sup.8 Day 11 Dog 5 F MSMA 10 Day 0 105804 10.sup.8 Blood G 140413 Months Day 3 3560 10.sup.8 Day 9 5314 10.sup.8 Day 11 183.7 10.sup.8 Dog 2 M LuMC5 3 Day 0 36630.6 10.sup.7 Blood M 151123 Months Day 3 1111.5 10.sup.7 Dog 3 M LuMC5 5 Day 0 102094.9 10.sup.7 Blood M 160301 Months Day 5 66.9 10.sup.7 Dog 5 M LuMC5 4 Day 0 62749.9 10.sup.7 Blood M 160308 Months Day 4 7.61 10.sup.7 Dog 3 M LuMC5 4 Day 0 6.66 10.sup.7 Blood M 160311 Months Day 4 Dog 5 M LuMC5 3 Day 0 6474.4 10.sup.7 Blood M 160324 Years Day 3 87.7 10.sup.7 Dog 8 F LuMC5 4 Day 0 107633.3 10.sup.7 Blood M 160326 Months Day 3 77.4 10.sup.7

(20) TABLE-US-00005 TABLE 2-4 Results of the clinical experiments on feline panleukopenia virus Animal Total Detecting Results & ID Age Gender Medicine Days point (ng/uL) Specimen Company Cat 3 F MSMA 12 Day 0 4240717 10.sup.8 Feces G 150323 Months Day 6 22391915 10.sup.8 (*c_) Day 15 711 10.sup.8 Day 21 14 10.sup.8 Cat 3 M MSMA 10 Day 0 130942 10.sup.8 Blood G 141128 Months Day 6 1088 10.sup.8 Day 7 25 10.sup.8 Day 8 79 10.sup.8 Cat 3 F MSMA 8 Day 0 143.67 10.sup.7 Blood G 140512 Years Day 7 Cat 3 F MSMA 12 Day 0 121.7 10.sup.7 Blood G 150529 Years Day 15 Cat 7 M LuMC5 7 Day 0 11.21 10.sup.7 Feces M 150826 Months Day 5 2.47 10.sup.7 (*c) Cat 8 M LuMC5 8 Day 0 40784.1 10.sup.7 Blood M 160104 Months Day 8 10.28 10.sup.7 Cat 8 F LuMC5 7 Day 0 4219.6 10.sup.7 Blood M 160108 Months Day 7 78.91 10.sup.7

(21) TABLE-US-00006 TABLE 2-5 Results of the clinical experiments on canine distemper virus Animal Total Detecting Results & ID Age Gender Medicine Days point (ng/uL) Specimen Company Dog 4 F MSMA 14 Day 0 1341 10.sup.8 Nasal G 140701 Months Day 4 334 10.sup.8 secretion Day 10 116 10.sup.8 (*c) Day 14 Dog 4 F MSMA 12 Day 0 24 10.sup.8 Nasal G 140703 Months Day 3 312 10.sup.8 secretion (*c) Day 10 61 10.sup.8 Day 12 Dog 1 F MSMA 11 Day 0 310 10.sup.8 Blood G 140808 Year Day 7 350 10.sup.8 Day 11 Dog 7 F MSMA 12 Day 0 350 10.sup.8 Blood G 140808-1 Years Day 8 264 10.sup.8 Day 12 Dog 12 M MSMA 3 Day 0 28 10.sup.8 Blood G 140617 Years Day 3

(22) TABLE-US-00007 TABLE 2-6 Results of the clinical experiments on feline coronavirus Animal Total Detecting Results & ID Age Gender Medicine Days point (ng/uL) Specimen Company Cat 5 F MSMA 8 Day 0 41733.8 10.sup.7 Blood G 150430 Years Day 8 Cat 4 M LuMC5 8 Day 0 5691.51 10.sup.7 Feces (*c) M 150826 Months Day 5 176.05 10.sup.7 Day 8 Cat 5 M LuMC5 5 Day 0 10589.77 10 Blood M 150903 Months Day 5 Cat 7 F LuMC5 8 Day 0 125.8 10.sup.7 Blood M 160110 Months Day 8 Ascites

(23) TABLE-US-00008 TABLE 2-7 Results of the clinical experiments on felid herpesvirus 1 Animal Total Detecting Results & ID Age Gender Medicine Days point (ng/uL) Specimen Company Cat 1 M MSMA 12 Day 0 1528214 10.sup.8 Blood G 150417 Year Day 4 214 10.sup.8 Day 13 Cat 5 M MSMA 10 Day 0 151.8 10.sup.7 Nasal G 150502 Months Day 10 3.75 10.sup.7 secretion (*c) Cat 3 F MSMA 9 Day 0 27380 10.sup. Blood G 150507 years Day 7 Cat 8 F MSMA 8 Day 0 2282.5 10.sup.7 Blood G 150516 Months Day 15 69.71 10.sup.7 Cat 8 F LuMC5 5 Day 0 326.1 10.sup.7 Blood M 151224 Months Day 5 Cat 11 F LuMC5 6 Day 0 8911.7 10.sup.7 Blood M 160315 Months Day 6 661.6 10.sup.7 *a: + refers to viruses were found in the detection. *b: refers to viruses were not found in the detection. *c: Not quantitative detection .circle-solid.: No pathogen infection or small amount of pathogens detected

(24) The administered period of MASA was 4 days to 14 days (average 9 days). The administered period of LuMC5 was 3 days to 9 days (average 5.7 days). When LuMC5 was administered via subcutaneous injection, intramuscular injection or intravenous injection, the fever in the subject was reduced within 3 hours to 5 hours, and metal condition of the subject was improved. The virus level in the specimens thereof was greatly reduced to slight amount within 2 days to 5 days, and reduced from the slight amount to the amount incapable to be detected within 5 days to 8 days. The infection syndromes were rapidly improved.

Embodiment 3

(25) In the present embodiment, LuMC5 was used to treat the mammalian with hemorrhagic venom infection and neurotoxic venom.

(26) Subject: Infected dogs and cates sent to Hei-Ming Veterinary Hospital, Taiwan

(27) Treatment: Intravenous injection or intramuscular injection, 0.4 cc/kg, once or twice a day

(28) The testing results s in the clinical experiments of the present embodiment are listed in the following Tables 3-1 to 3-3.

(29) TABLE-US-00009 TABLE 3-1 Results of the clinical experiments on the subject infected with hemorrhagic venom infection of Trimeresurus mucrosquamatus Animal Time to Administered Syndromes before treatment & ID Age Gender be treated times Condition after treatment Dog 3 M 2 hours 1 Depression, lifeless, sticky saliva, 130420 years after and facial and head swelling infection Depression and lifeless were relieved, 1 hours after treatment. Facial and head swelling were relieved, 12 hours after treatment. Dog 6 F 2 hours 2 Facial swelling, depression, lifeless, 140405 years after and mouth and tongue blood stasis infection Depression and lifeless were relieved, 30 minutes after treatment. Facial swelling and mouth and tongue blood stasis were relieved, 24 hours after treatment. Cat 1 F 1 hours 2 Shock, unconsciousness, tachypnoea, 140320 year after transparent and jelly-like saliva, and infection hematuria Shock, unconsciousness, tachypnoea, and transparent and jelly-like saliva were relieved, 30 minutes after treatment. Hematuria was relieved and the subject became healthy, 50 minutes after treatment. Dog 3 M 26 hours 2 Depression, lifeless, sticky saliva, and 140724 years after chin swelling infection Depression, lifeless, and sticky saliva were relieved, 50 minutes after treatment. Chin swelling was relieved, 12 hours after treatment. Dog 4 M 4 hours 1 Bleed and depression 140707 years after Bleed caused by snakebite was relieved, infection 10 minutes after treatment. Depression was relieved, 30 minutes after treatment. Dog 4 M 4 hours 1 Bleed and depression 140707-1 years after Bleed caused by snakebite was relieved, infection 10 minutes after treatment. Depression was relieved, 30 minutes after treatment. Dog 7 M 16 hours 2 Depression, lifeless, sticky saliva, 141028 years after and facial and chin swelling infection Depression and lifeless were relieved, 30 minutes after treatment. Facial and chin swelling was relieved, 12 hours after treatment.

(30) TABLE-US-00010 TABLE 3-2 Results of the clinical experiments on the subject infected with hemorrhagic venom infection of Viridovipera stejnegeri Animal Time to Administered Syndromes before treatment & ID Age Gender be treated times Condition after treatment Dog 1 M 14 hours 3 Depression, lifeless, sticky saliva, 140412 year after and facial and chin swelling infection Depression and lifeless were relieved, 2 hours after treatment. Facial and chin swelling was relieved, 24 hours after treatment.

(31) TABLE-US-00011 TABLE 3-3 Results of the clinical experiments on the subject infected with neurotoxic venom infection of Naja Animal Time to Administered Syndromes before treatment & ID Age Gender be treated times Condition after treatment Dog 6 M 2 hours 2 Depression, lifeless, sticky saliva, 140426 years after and facial and chin swelling infection Depression and lifeless were relieved, 30 minutes after treatment. Facial and chin swelling was relieved, 12 hours after treatment.

(32) From the results shown in the present embodiment, when LuMC5 was administered into the infected subject via subcutaneous injection, intramuscular injection or intravenous injection, the nose bleeding and sticky saliva can be relieved within 10 minutes, the depression can be relieved within 30 minutes, the hematuria can be relieved within 50 minutes, and the swelling can be relieved and the treated subject become healthy within 1 hours to 36 hours. Hence, by using LuMC5 of the present disclosure, the infection syndromes can be controlled. Compared to the conventional method by naturalizing toxins with serum formulation, in which antibodies are formed to identify and neutralize toxin proteins after 7 days to 14 days and the swelling syndrome is relieved after 7 days, LuMC5 of the present disclosure can relieve the syndromes in a short time and has superior treatment efficacy.

Embodiment 4

(33) In the present embodiment, LuMC5 was used to treat the mammalian that the mosquito, bee, scorpion, spider, centipede, ant, and staphylinidae bitten.

(34) Subject: Dog bit by bees, sent to Hei-Ming Veterinary Hospital, Taiwan, and having bodyweight of 27 Kg

(35) Treatment: Intravenous injection or intramuscular injection, 0.4 cc/kg, twice a day

(36) The result is shown in the following Table 4-1.

(37) TABLE-US-00012 TABLE 4-1 Animal Time to Administered Syndromes before treatment & ID Age Gender be treated times Condition after treatment Dog 14 F 2 hours 2 Depression, lifeless, and 140414 years after facial and head swelling infection Depression and lifeless were relieved, 30 minutes after treatment Facial and head swelling was relieved, 12 hours after treatment.

(38) The treatment results show that the mental condition of the treated subject was improved, the appetite thereof was recovered, and the activity thereof such as tail shaking turned into normal, after treatment.

(39) Subject: Human bit by a mosquito, bodyweight: 79 Kg

(40) Treatment: Applying LuMC5 at the mosquito biting region, twice a day

(41) The result is shown in the following Table 4-2.

(42) TABLE-US-00013 TABLE 4-2 Animal Time to Administered Syndromes before treatment & ID Age Gender be treated times Condition after treatment Dog 55 F 3 minutes 2 Swelling and itch at the 160415 years after biting region on the skin infection Itch was relieved, 3-20 minutes after applying medicine. Swelling was relieved, 1-3 hours after applying medicine.

(43) From the results shown in the present embodiment, LuMC5 can effectively relieve the itch and selling on the skin caused by the insects. If the subject is bit by the insects, the glycosylation of the virus can be inhibited when LuMC5 is applied immediately. In addition, the syndromes of venom allergy, fever and lifeless can be relieved by administering LuMC5 via injection. The swelling at the skin region bit by the insects with pathogens can be relieved by applying ointment or solution at the affected region. Therefore, Dengue virus, West Nile virus and Zika virus infection to human and animals can be effectively relieved and prevented by using LuMC5 of the present disclosure.

Embodiment 5

(44) In the present embodiment, LuMC5 was used to treat cataract and retinopathy caused by abnormal eye cells in mammalian.

(45) Subject: Infected dogs and cates sent to Hei-Ming Veterinary Hospital, Taiwan

(46) Treatment: Solution containing 1-2 mg/cc of phenol red and 3-4 mg/cc of monosodium methanearsonate, one drop per time, twice or three times a day

(47) The result is shown in the following Tables 5-1 and 5-2.

(48) TABLE-US-00014 TABLE 5-1 Results of the clinical experiments on Pomeranian with cataract Animal Syndromes before treatment & ID Age Gender Total days Treatment Condition after treatment Dog 18 F 46 One drop Mature cataract, spin around while walking, 151013 years per time, incapable of walking straight, hitting the three times wall, and recognizing surrounding by a day smelling, auditory sense and living experience The clouding of the lens in the eye reduced. The treated subject can walk straight without hitting the wall.

(49) TABLE-US-00015 TABLE 5-2 Results of the clinical experiments on Golden Retriever with cataract Animal Syndromes before treatment & ID Age Gender Total days Treatment Condition after treatment Dog 10 M 29 One drop The subject cannot see the object at 160128 years per time, the long distance during daytime. twice a day Visually impaired during the night. The clouding of the lens in the eye reduced. The subject's day and night vision was recovered.

Embodiment 6

(50) In the present embodiment, LuMC5 was used to treat metabolism-related diseases (including pancreatitis, kidney inflammation, hepatitis, and cholangitis) occurred in mammalian.

(51) Subject: Infected dogs and cates sent to Hei-Ming Veterinary Hospital, Taiwan

(52) Treatment: Intravenous injection or intramuscular injection, 0.4 cc/kg, once a day

(53) Testing company: LEZEN Reference Lab

(54) The result is shown in the following Tables 6-1 to 6-4.

(55) TABLE-US-00016 TABLE 6-1 Animal Body Syndromes before treatment & ID Age Gender Total days weight Condition after-treatment Dog 15 M 11 20 Kg Anorexia for 4-5 days, urine in dark 160714 years brown, weakness, and dehydration Appetite and activity recovered

(56) TABLE-US-00017 TABLE 6-2 Blood test data of Dog 160714 during the treatment PT:ALT Alkaline-p BUN Creatinine Amylase (liver) (gall) (uremia) (Kidney) (pancreas) Day 1 87 87 94.2 3.44 1447 Day 19 24 50 60 2.78 1225 Reference 14-40 32-91 8-20 0.64-1.27 S: 28-1200 U/L U/L mg/dL mg/dL U/L

(57) TABLE-US-00018 TABLE 6-3 Animal Total Body Syndromes before treatment & ID Age Gender days weight Condition after treatment Dog 15 F 3 28 Kg Anorexia for 1-2 days, vomit, 160805 years weakness, incapable of standing Capable of standing at Day 2, eating small amount of food, and walking slowly at Day 3

(58) TABLE-US-00019 TABLE 6-4 Blood test data of Dog 160805 during the treatment BUN Creatinine Amylase Item (uremia) (Kidney) (pancreas) 2016 Aug. 5 31.5 2.46 1251 2016 Aug. 8 21.5 1.19 931 Reference 8-20 0.64-1.27 S: 28-1200 mg/dL mg/dL U/L

(59) From the results shown in Embodiments 5 and 6, a solution made from LuMC5 can be used to treat diseases caused by abnormal eye cells (such as cataract and retinopathy) by administering the solution at the predetermined time with a predetermined amount. When treating diseases caused by abnormal brain cells (including Alzheimer's Disease and Parkinson's disease), LuMC5 can be formulated into a paste or ointment and administered at the predetermined time with a predetermined amount. The skin absorbs the active ingredients, and the active ingredients can enter into the brain via the blood in the carotid artery to accomplish the purpose of preventing disease. When treating metabolism related diseases (such as pancreatitis, kidney inflammation, hepatitis, as cholangitis), LuMC5 can be administered via subcutaneous injection, intramuscular injection or intravenous injection for 5 to 7 days, and the swelling and inflammation can be effectively relieved.

Embodiment 7

(60) When treating prion infection (including bovine spongiform encephalopathy, scrapie, and creutzfeldt-Jakob disease), LuMC5 can be formulated into a paste or ointment and administered at the predetermined time with a predetermined amount. The skin absorbs the active ingredients, and the active ingredients can enter into the brain via the blood in the carotid artery to accomplish the purpose of preventing disease.

(61) Although the present invention has been explained in relation to its preferred embodiment, it is to be understood that many other possible modifications and variations can be made without departing from the spirit and scope of the invention as hereinafter claimed.

Embodiment 8

(62) The following experiments were conducted at the Center for Infectious Disease and Cancer Research, Kaohsiung Medical University, in Taiwan.

(63) Materials and Methods

(64) 1. Cell Model: CD4.sup.+ molecules-expressing cell line: MAGIC-5 cell

(65) 2. HIV Resource: HIV infectious clone: HIV-1 CRF07_BC HIV replication activity will be determined by blue-forming (BFU) assay.

(66) 3. Antiretroviral Drugs: LuMC5 (Phenol red 0.2% and Monosodium methanearsonate 0.4%)

(67) 4. AlamarBlue: AlamarBlue assay (AbD Serotec) was used to evaluate cell viability and cell proliferation according to the manufacturer's instruction. In brief, DMEM and MAGIC-5 cells (4.510.sup.3 cells/well) were seeded and treated with or without LuMC5 (25 dilution to 800 dilution). A DDW treated group was conducted as a solvent control. After 72 hours of incubation at 37 C., 10% alamarBlue was added and further incubated for 4 hours at 37 C. to assess the cytotoxic effects of the tested drugs. The result of cell viability assay has normalized by solvent control. After 4 hours of AlamarBlue addition fluorescence of the reduced AlamarBlue was recorded in a microplate reader (Synergy HT, Biotek Instruments, Winooski, USA).

(68) 5. Blue-Form Unit (BFU) Assay (MAGIC-5 HIV-1 Infectivity Assay): In order to estimate the drug susceptibility of CRF07_BC with LuMC5, the HIV-1 infectivity would be assessed by blue-form unit (BFU) assay. BFU assay is an in-house phenotypic assay based on the property of MAGIC-5 cells that BFU could be formed in cells after HIV-1 infection. MAGIC-5 cells were counted and dispensed into a 96-well tissue culture plate at 4.510.sup.3 cells/well with growth medium (DMEM with 10% FBS, NEAA, P/S and L-glutamine) 12-hour before infection. After cells were attached, 100 BFUs of virus stock were added into wells in triplicate. The supernatants were discarded after 4-hour 37 C. incubation, and the wells were refilled by virus-free fresh growth medium with or without serial-diluted LuMC5 (50-800). A 0.84 g/m of Lamivudine (3TC)one of the common and regular HAART drugs-treated group was conducted as a positive control. Cells were assayed for infection by staining for -galactosidase expression at 48-hour post-infection. Culture medium was removed, and fixing solution (0.1% formaldehyde and 0.02% glutaraldehyde in PBS) was added to each well. Cells were fixed at room temperature for 5 min, and 100 l of staining solution (4 mM potassium ferrocyanide, 4 mM potassium ferricyanide, 2 mM magnesium chloride, and 400 g/ml 5-bromo-4-chloro-3-indolyl--D-galactopyranoside [X-gal] in PBS) was added to each well for 1-4 hours. The number of blue-stained cells were counted and be considered as blue cell-forming units (BFUs). The results of BFU assay have been normalized by the cell survivability data from AlamarBlue assay that described above to eliminate the cell toxicity effects from LuMC5. Student's t-test has been calculated for the statistics examination.
Objectives: 1. To conduct a cell viability assay to assess the cytotoxicity of LuMC5 in MAGIC-5 cell model. 2. To determine the viral inhibitory effects of LuMC5 on HIV-1 CRF07_BC via MAGIC-5 cell-based HIV infectivity assay.
Results:

(69) 1). MAGIC-5 Cell Cytotoxicity Report of LuMC5

(70) The result of AlamarBlue assay has shown that, the cell viability of MAGIC-5 was decreased to 63.9% (1.83) after 25-folds LuMC5 treatment. In the 50-folds treated group, the cell viability was reached to 82.4% (2.24) (FIG. 2). FIG. 2 is a figure showing MAGIC-5 Cell viability assay of LuMC5, wherein the H.sub.2O treatment group has considered a solvent control; all value of cell viability has normalized by the no drug treatment control, MeanSD. The 50-folds dilution was chosen and actually is the closest concentration to LD.sub.20, to be the base-line treatment dose during drug susceptibility experiment.

(71) 2). LuMC5 Drug Susceptibility Via BFUs HIV-1 Infectivity Assay

(72) The result of CRF07_BC drug susceptibility assay has shown that the viral inhibition ability of LuMC5 with HIV-1 CRF07_BC is significant and showed dose-dependent effects (50, 100, 200, 400, 800 of LuMC5 inhibit 39.3%, 37.6%, 28.8%, 31.1% and 25.5% of HIV-1 CRF07_BC infectivity respectively. Student's t-test has been conducted for the comparison between ND and other different LuMC5 treatment groups, p<0.05) (FIG. 3). FIG. 3 is a figure showing drug susceptibility assay of LuMC5 treated in HIV-1 CRF07_BC infection model, wherein all viral infectivity assay result has normalized by cell viability data from AlamarBlue assay. The result of no drug treatment group (ND) was be considered as 100% of virus infectivity

(73) In the above experiments, it was found that LuMC5 (0.2% phenol red and 0.4% monosodium methanearsonate) at 50 dilution could inhibit 39.3% of HIV-1 CRF07_BC infectivity in MAGIC-5 cell model and showed dose-dependent effects, i.e. the higher dosage, the higher inhibition rate. Even the dosage after 800 dilution (i.e. 2.5 ppm phenol red and 5 ppm monosodium methanearsonate) still have inhibition rate of 25.5%. These results prove that LuMC5 is able to inhibit human immunodeficiency virus (HIV) to quite meaningful level.

(74) Usually, the patients are affected by HIV majorly through sexual intercourse, blood transfusion, or wound contact. Thus, LuMC5 can be made into the form of ointment or lotion, which can be applied onto or dosed into a wound ointment, a condom, a gauze, or a medical tool, such as a syringe, a medical tube, or a surgical tool for preventing people from being affected by HIV.