Cyclic poly L-lactic acid

Abstract

A cyclic poly L-lactic acid composition having a pH of 6 to 7 by the polymer itself.

Claims

1. A cyclic poly L-lactic acid composition having a pH of 6 to 7 by the polymer itself, wherein the cyclic poly L-lactic acid has a cyclic polymerization degree (n) of 5 to 30, and wherein the cyclic poly L-lactic acid having the cyclic polymerization degree (n) of 5 to 10 shows an ionic strength of 50% or more.

2. The cyclic poly L-lactic acid composition according to claim 1, further comprising a chain poly L-lactic acid having a chain polymerization degree (n) of 2 to 30.

3. A pharmaceutical product in which the cyclic poly L-lactic acid composition according to claim 1 is compounded.

4. A pharmaceutical product in which the cyclic poly L-lactic acid composition according to claim 2 is compounded.

Description

BRIEF EXPLANATION OF DRAWINGS

(1) FIG. 1

(2) The mass spectrum of Patent Document 1 is shown.

(3) FIG. 2

(4) The mass spectrum (MALDI) of the sample is shown.

(5) FIG. 3

(6) The mass spectrum (MALDI, enlarged) of the sample is shown.

(7) FIG. 4

(8) The analysis result by polytools is shown.

(9) FIG. 5

(10) The mass spectrum (ESI+) of the sample is shown.

(11) FIG. 6

(12) The mass spectrum (ESI+, enlarged) of the sample is shown.

ASPECTS OF CARRYING OUT THE INVENTION

(13) In the production method of the cyclic poly L-lactic acid, L-lactic acid solution is put into a 50 L mantle heater in a production machine tank, heat treatment is performed at a temperature of 100 C. for several hours. The pressure is then gradually reduced by a vacuum pump, and nitrogen gas is injected directly on a side area of the tank so as to stir the inside of the tank, control the pressure and discharge the generating steam. The temperature during heating is increased from 100 C. to reach the pressure of (minus) 525 mmHg and temperature of 170 C. after 7 hours. After the pressure is further decreased to (minus) 600 mmHg, the pressure is raised and lowered several times between (minus) 600 mmHg and (minus) 225 mmHg at a temperature of from 170 C. to 177 C. to generate a large amount of steam. As the temperature is raised in several hours, the pressure is decreased to the pressure of (minus) 975 mmHg and temperature of 193 C. The decompression is then stopped while maintaining the temperature at 193 C. After the pressure is shifted to 760 mmHg, heat-treatment is continued for several hours. The substance is then taken out, and cooled and solidified at 60 C. to give a high purity substance of the cyclic poly L-lactic acid with no acidity, colorless, and transparency.

(14) In order to examine the polymerization degree of the cyclic poly L-lactic acid, matrix-assisted laser desorption/ionization/mass spectrometry (MALDI/MS) and electrospray ionization/mass spectrometry (ESI/MS) were performed. As a result, although a monomer unit of L-lactic acid (C3H4O2) with molecular weight distribution of m/z 72 difference was confirmed, it was also shown that the cyclic poly L-lactic acid (degree of polymerization: 5 to 30) and the chain or linear (straight) poly L-lactic acid (degree of polymerization: 2 to 30) coexisted.

(15) [Chemical Formula 1]

(16) The chemical formulae of the cyclic poly L-lactic acid and the chain poly L-lactic acid are shown.

(17) ##STR00001##

(18) Cyclic poly L-lactic acid (degree of polymerization: 5 to 30)

(19) ##STR00002##

(20) Chain poly L-lactic acid (degree of polymerization: 2 to 30)

(21) Analytical method of the cyclic poly L-lactic acid of the present invention

(22) Analytical apparatus

(23) Matrix-Assisted Laser Desorption/Ionization/Mass Spectrometry (MALDI/MS)

(24) Apparatus: rapid FLEXX TOF/TOF type manufactured by Bruker Daltonics

(25) Measurement mode: Reflector/Positive mode

(26) Measurement mass range: m/Z 100-4000

(27) Electrospray Ionization/Mass Spectrometry (ESI/MS)

(28) <Liquid Chromatograph Unit>

(29) Apparatus: ACQUITY UPLC type manufactured by Waters

(30) Mobile phase: 0.1% formic acid aqueous solution/acetonitrile=1/1

(31) Flow rate: 0.2 ml/min

(32) <Mass Analyzer>

(33) Device: Synapt G2-S type manufactured by Waters

(34) Measurement mode: Positive mode

(35) Measurement mass range: m/z 50 to 2000

(36) Analysis Method

(37) MALDI/MS: A sample was dissolved in tetrahydrofuran, mixed with matrix (DHB) solution, and subjected to the analysis.

(38) ESI/MS: A sample was dissolved in tetrahydrofuran and subjected to the analysis.

EXAMPLE

(39) L-lactic acid solution (40 kg) with 90% purity was put into the 50 L mantle heater in the production machine tank, heat treatment was performed at a temperature of 100 C. for several hours. The pressure was then gradually reduced by the vacuum pump, and nitrogen gas was injected directly on the side area of the tank at 1.3 L/min so as to stir the inside of the tank, control the pressure and discharge the generating steam. The generating steam and nitrogen gas were cooled while being passed through a cooling apparatus, so that the steam was taken out as liquid and stored in a 20 L tank so as to be separated from nitrogen gas. The temperature during heating is increased from 100 C. while decompressing gradually by (minus) 75 mmHg per 60 min to reach the pressure of (minus) 525 mmHg and temperature of 170 C. after 7 hours. After the pressure was further decreased by (minus) 75 mmHg per 30 min to (minus) 600 mmHg, the pressure was raised and lowered several times between (minus) 600 mmHg and (minus) 225 mmHg at a temperature of from 170 C. to 177 C. to generate a large amount of steam. The pressure was further decreased by (minus) 75 mmHg per 10 min and the temperature was according increased to reach the pressure of (minus) 975 mmHg and temperature of 193 C. The decompression and the injection of the nitrogen gas were stopped while maintaining the temperature at 193 C. After the pressure was shifted to 760 mmHg by introducing oxygen to generate the steam, heat-treatment was continued for several hours. The substance was then taken out, cooled and solidified at 60 C. to give a high purity substance (25 kg) of the cyclic poly L-lactic acid with no acidity, colorless, and transparency.

(40) Explanation of the Substances

(41) TABLE-US-00001 Cyclic poly L-lactic acid of the present Conventional cyclic L-lactic Name invention poly L-lactic acid acid Lactide Poly L-lactic acid Molecular Formula (C.sub.3H.sub.4O.sub.2)n (C.sub.3H.sub.4O.sub.2)n C.sub.3H.sub.6O.sub.3 C.sub.6H.sub.8O.sub.4 (C.sub.3H.sub.4O.sub.2)n Molecular Weight Unit m/z 72 Unit m/z 72 90.08 144.1 90,000~280,000 low molecular low molecular high molecular Cyclic polymerization n = 5-30 n = 3-19 n = 1 n = 2 degree n = 9-19 Chain polymerization m = 2-30 m = 5-23 degree Cyclic polymerization Ionic strength of Ionic strength of degree n = 5-10 50% or more 15% or less pH 6-7 2-3 1-2 Acidity no yes yes Neutralizing agent not necessary necessary Sweetener not necessary necessary

(42) Analysis Results and Discussion of the Cyclic Poly L-Lactic Acid of the Present Invention

(43) MALDI/MS

(44) MALDI/MS was performed to examine the degree of polymerization of the cyclic poly L-lactic acid. FIGS. 2 and 3 show the mass spectrum (MALDI) of the sample. Ion groups having a distribution of m/z 72 difference, which corresponds to a monomer unit of L-lactic acid, were confirmed. FIG. 4 shows the results of analysis by a polymer analysis software (polytools; manufactured by Bruker Daltonics). When the repeating unit is set to m/z 72 for poly lactic acid (PLA), a terminal group of the first (red) and fifth (green) ion groups (presumed as sodium ion-addition molecule (M+Na).sup.+, and potassium ion-addition molecule (M+K).sup.+, respectively) detected with a high intensity was presumed to be OH and H, respectively, suggesting that they were a chain (straight) structure. The eighth (blue) and thirteenth (blue-green) ion groups were estimated to be protonated molecules (M+H).sup.+ and (M+Na).sup.+ of the cyclic poly L-lactic acid, respectively. The ions with the thus detected degree of polymerization of 5-30 are shown in FIG. 3. The end groups of other ions were not identified.

(45) TABLE-US-00002 TABLE 1 Results of Composition Estimation with the accurate mass Elemental Composition Report Single Mass Analysis Tolerance = 3.0 mDa/DBE: min = 1.5, max = 50.0 Element prediction: Off Number of isotope peaks used for i-FIT = 3 Mass Difference m/z 72 Monoisotopic Mass, Odd and Even Electron Ions 8 formula(e) evaluated with 1 results within limits (all results (up to 1000) for each mass) Elements Used: C: 0-100 H: 0-100 O: 0-20 161201_01 213 (0 810) Cm (208:216-114:137) Mass Calc. Mass mDa PPM DBE Formula 72.0214 72.0211 0.3 4.2 2.0 C3 H4 O2 Monoisotopic Mass, Even Electron Ions 106 formula(e) evaluated with 2 results within limits (all results (up to 1000) for each mass) Elements Used: C: 0-100 H: 0-100 O: 0-20 161201_01 213 (0.810) Cm (208:216-114:137) Mass Calc. Mass mDa PPM DBE Conf (%) Formula (i) Blue 505.1567 505.1557 1.0 2.0 7.5 100.00 C21 H29 O14 505.1592 2.5 4.9 29.5 0.00 C39 H21 O Monoisotopic Mass, Even Electron Ions 220 formula(e) evaluated with 2 results within limits (all results (up to 1000) for each mass) Elements Used: C: 0-100 H: 0-100 N: 0-1 O: 0-20 161201_01 213 (0.810) Cm (208:216-114:137) Mass Calc. Mass mDa PPM DBE Conf (%) Formula (II) Yellow 522.1830 522.1858 2.8 5.4 28.5 58.26 C39 H24 N O 522.1823 0.7 1.3 6.5 41.74 C21 H32 N O14 Monoisotopic Mass, Even Electron Ions 114 formula(e) evaluated with 2 results within limits (all results (up to 1000) for each mass) Elements Used: C: 0-100 H: 0-100 O: 0-20 161201_01 213 (0.810) Cm (208:216-114:137) Mass Calc. Mass mDa PPM DBE Conf (%) Formula (III) Red 523.1682 523.1663 1.9 3.6 6.5 99.99 C21 H31 O15 523.1698 1.6 3.1 28.5 0.01 C39 H23 O2 Monoisotopic Mass, Even Electron Ions 229 formula(e) evaluated with 2 results within limits (all results (up to 1000) for each mass) Elements Used: C: 0-100 H: 0-100 N: 0-1 O: 0-20 161201_01 213 (0.810) Cm (208:216-114:137) Mass Calc. Mass mDa PPM DBE Conf (%) Formula (IV) Green 540.1937 540.1928 0.9 1.7 5.5 100.00 C21 H34 N O15 540.1964 2.7 5.0 27.5 0.00 C39 H26 N O2 0 ESI/MS In order to clarify the terminal structure of the poly L-lactic acid, composition estimation was performed with ESI/MS (high resolution). The mass spectrum (ESI+) of the samples is shown in FIG. 5 and FIG. 6, and the results of composition estimation with the accurate mass are shown in Table 1. From the mass spectrum, the ion groups having a repeating unit of m/z 72 were detected, which was estimated with the accurate mass to be a lactic acid unit (C3H4O2). In addition, the composition estimation was performed for the representative ions (m/z 505, 522, 523 and 540) of the ion groups shown by (I) blue, (II) yellow, (III) red and (IV) green circles. As a result, the (I) blue and (II) yellow circles were estimated to be (M + H).sup.+ and an ammonium ion-adduct molecule (M + NH4).sup.+, respectively, of the cyclic poly L-lactic acid, and the (III) red and (IV) green circles were estimated to be (M + H).sup.+ and (M + NH4).sup.+, respectively, of the chain poly L-lactic lactic acid. In addition, ion groups having a degree of polymerization of 2 (m/z 145) or more were detected for the cyclic poly L-lactic acid.

(46) Analytical Results of the Cyclic Poly L-Lactic Acid According to the Invention: Summary

(47) The matrix-assisted laser desorption/ionization/mass spectrometry (MALDI/MS) was performed to determine the degree of polymerization of the cyclic poly L-lactic acid. As a result, the ion groups were detected, which had the distribution of m/z 72 difference corresponding to the monomer unit of L-lactic acid. The results by the software suggested the presence of not only the cyclic poly L-lactic acid (degree of polymerization: 5 to 30) but also the chain poly L-lactic acid. Therefore, in order to investigate in more detail the terminals of the poly L-lactic acid, high resolution measurement with ESI/MS was performed. As a result, it was also presumed that the ion groups detected with ESI/MS had the cyclic (degree of polymerization: 2 or more) and chain structures.

(48) The present invention also relates to a pharmaceutical product in which the thus newly prepared cyclic poly L-lactic acid (composition) is compounded. As described in Patent Document 3 above, it is known that the cyclic poly L-lactic acid has various physiologically actions. Therefore, based on such actions the pharmaceutical product of the present invention has various medical uses such as, for example, a metabolic regulator, a metabolic inhibitor, an immune activator (for example, activator of immune cells such as NK cells), an inhibitor of LDH activity of a cancer cell, an inhibitor/suppressor of the growth of a cancer cell, and a cancer therapeutic agent. It may be administrated orally as oral powder, tablets, capsules, and parenterally drops, injections, poultices, and the like.

(49) An amount of Ingestion of the oral powder of the pharmaceutical product comprising the cyclic poly L-lactic acid composition of the present invention is preferably 6 to 15 g per day, and the effect can be observed in one to three months.

(50) The effects of the pharmaceutical product according to the present invention in cases.

(51) Case 1: Gastric Cancer and Lung Cancer Metastasis

(52) A 70-year-old man having gastric cancer diagnosed by a medical checkup but not treated after that, and then had a medical examination again so that lung metastatic cancer was diagnosed. After daily intake of 15 g of the cyclic poly L-lactic acid powder of the present invention for 60 day, PET examination was performed to show that the lung cancer and the gastric cancer had disappeared.

(53) Case 2: Skin Cancer

(54) A 60-year-old woman, with brownish tumor-like breast that had been blackened daily, and shades in lungs. After daily intake of 15 g of the cyclic poly L-lactic acid powder of the present invention for 6 months, a part of the surface of the skin was removed, the shades in lungs had disappeared.

(55) Case 3: Lung Cancer

(56) A 62-year-old man had developed lung cancer, which was unable to treat according to the diagnosis at a hospital. After daily intake of 15 g of the cyclic polyL-lactic acid powder of the present invention for 2 months, X-ray examination revealed that the lung cancer had disappeared.

(57) Case 4: Cervical Cancer

(58) A 45-year-old woman with cervical cancer according to diagnosis was going to undergo surgery 3 months later. After daily intake of 6 g of the cyclic poly L-lactic acid powder daily according to the present invention for two months, an examination before the surgery revealed that the cancer had disappeared.

(59) Case 5: Breast Cancer

(60) A 72-year-old woman felt lumps on her chest, which was diagnosed at a hospital the next day as 20 mm breast cancer. After daily intake of 10 g of the cyclic poly L-lactic acid powder of the present invention for 1 year, MRI examination revealed that the breast cancer had disappeared.

(61) Case 6: Colorectal Cancer

(62) A 72-year-old man was examined to have a colorectal polyp in a specific health checkup, and 9 polyps were removed under surgery. After daily intake of 6 g of the cyclic poly L-lactic acid powder of the present invention for 1 year, one polyp was found, excised and diagnosed as a malignant tumor. No recurrence has occurred thereafter.

(63) Case 7: Gastric Cancer

(64) A 74-year-old man with gastric cancer according to diagnosis after an examination at a hospital. After two thirds of the stomach had been removed, he was recommended that he should immediately take surgery as the liver had also a shade, and should also take anti-cancer drug treatment. After daily intake of 10 g of the cyclic poly L-lactic acid powder of the present invention for 3 months, an endoscopic examination performed after 6 months revealed that the stomach cancer had disappeared.

(65) Case 8: Gastric Cancer

(66) A 64-year-old man was found to have four tumors in the stomach with a gastroscope examination under a health examination. After daily intake of 10 g of the cyclic poly L-lactic acid powder according to the present invention for 2 months, it was revealed that three tumors had disappeared and one tumor had been reduced, so that surgery could be avoided.

(67) Case 9: Pancreatic Cancer

(68) A 63-year-old woman was unwell and underwent a thorough examination at a hospital, so that pancreatic cancer was diagnosed.

(69) Due to the side effect of an anti-cancer drug treatment, she had become unable to eat, and the physical fitness had accordingly declined. After daily intake of 10 g of the cyclic poly L-lactic acid powder for 2 months, MRI examination revealed that the pancreatic cancer had become smaller.

(70) Case 10: Liver Cancer

(71) A 65-year-old man with liver cancer diagnosed at a hospital had taken daily 10 g of the cyclic poly L-lactic acid powder of the present invention for 3 months. The results of MRI examination revealed that the liver cancer had disappeared.

(72) Case 11: Hemangioma

(73) A 33-year-old woman had bleeding from a tumor in two thirds of the left nose. After daily intake of 6 g of the cyclic poly L-lactic acid powder of the present invention for 2 months, the tumor had disappeared and surgery had become unnecessary according to diagnosis.

(74) Case 12: Prostate Cancer

(75) A 64-year-old man with hematuria had an examination in a urology department so that prostate cancer was diagnosed. A PSA value was 1519. It was said that it was difficult to treat due to decrease in physical fitness, reduction in immunity, loss of appetite, and gait difficulty. After daily intake of 10 g of the cyclic poly L-lactic acid powder of the present invention for 3 months, a re-examination showed a PSA value had gone down to 7.6.

(76) Case 13: Colorectal Cancer

(77) A 70-year-old man had taken the colorectal cancer surgery to remove large intestine of about 15 cm and taken medication to suppress metastasis. But, the examination of the large intestine done 2 years after the operation revealed two polyps in the intestine so that an operation was planned to be done three months later. However, after daily intake of 6 g of the cyclic poly L-lactic acid powder of the present invention for three months before the operation, an endoscopic examination revealed only one polyp remaining, which had been shrunk, and the other one had disappeared.

(78) Case 14: Regressive Rheumatism

(79) A 58-year-old man with continued physical abnormality and pain run throughout the body took examination at a hospital. The diagnosis result identified rheumatic rheumatism. Taking prescribed medicine had to be stopped due to the occurrence of abnormality in liver function. After daily intake of 6 g of the cyclic poly L-lactic acid of the present invention for 2 months, the pain in the whole body had been alleviated and blood test values had been also decreased.

(80) Case 15: Cervical Cancer

(81) A 60-year-old woman, after having undergone cervical cancer excision surgery and taken radiotherapy several times, suffered from metastasis to the vagina, which was not possible to treat. After daily intake of 10 g of the present invention cyclic poly L-lactic acid powder of the present invention for 3 months, the diagnosis by a hospital said that there had been no abnormality.

(82) Case 16: Lung Cancer Stage 4

(83) A 65-year-old woman having suffered from lung cancer was told after diagnosis that it was inoperable. After daily intake of 15 g of the cyclic poly L-lactic acid powder of the present invention for 3 months, MRI examination revealed that the lung cancer had become smaller.

(84) Case 17: Small-Cell Lung Cancer

(85) A 70-year-old man with small-cell lung cancer was admitted to a hospital, underwent surgery, and received anticancer drug treatment. Due to its metastasis to the brain, he had taken radiation therapy, and continued anti-cancer drug treatment. The anti-cancer drug treatment was then suspended. After daily intake of 10 g of the cyclic poly L-lactic acid powder of the present invention for 3 months, she had lived the same life as before as her physical condition had returned and.

(86) Case 18: Metastasis of Uterine Cancer to the Whole Body

(87) A 60-year-old woman was hospitalized with uterus cancer, but according to diagnosis, the cancer had metastasized to the stomach and the whole body and it could not be treated. While she had cleaned up her belongings, she had taken daily 15 g of the cyclic poly L-lactic acid powder of the present invention for 6 months. The body condition had become healthy, and it was told by the doctor that the cancer cells metastasized to the whole body had become smaller according to MRI examination by the hospital.

(88) Case 19: Breast Cancer and Nasal Hemangioma

(89) A 63-year-old woman had suffered from breast cancer recurrence and nasal hemangioma according to diagnosis. After daily intake of 10 g of the cyclic poly L-lactic acid powder of the present invention for 2 months, the breast cancer and the nasal hemangioma had shrunk according to the examination and diagnosis at the hospital.

(90) Case 20: Spinal Canal Stenosis:

(91) A 57-year-old man with sudden back pain suffered walking and unable to turn over was hospitalized. As a result of diagnosis of spinal stenosis, it was recommended that he should take surgery. After daily intake of 10 g of the cyclic poly L-lactic acid powder of the present invention for one month, the pain had suddenly disappeared and it had become possible to walk and turn over, so that surgery could be avoided.