JP2005529903A - Use of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) for secondary brain disease events such as stroke - Google Patents

Abstract

An object of the present invention is to provide an effective and safe method for helping to prevent cerebral disease events, particularly cerebral infarction, in patients with symptoms of atherosclerosis of arteries supplying blood to the brain. And
The use of n-3 PUFA EPA and / or DHA in the preparation of an oral drug for preventing brain damage in a patient exhibiting symptoms of atherosclerosis of an artery supplying blood to the brain To do.

Description

  The present invention relates to n-3 PUFA eicosapentaenoic acid (EPA) and / or docosahexaenoic acid in the prevention of secondary brain disease events, particularly stroke, in patients exhibiting symptoms of atherosclerosis of the arteries supplying blood to the brain Relates to the use of (DHA).

  Consumption of fish or long-chain n-3 polyunsaturated fatty acids (PUFA), especially oily fish and fish oils, such as EPA and DHA, prevents cardiovascular disease in Westerners There is a lot of epidemiological evidence. Long-chain n-3 PUFA reduces triacylglycerol (TAG) concentration in fasting plasma and reduces postprandial lipemic response. Animal models have already demonstrated that edible fish oil reduces atherosclerosis, but this is due to reduced lipids, reduced growth factor production, reduced inflammation, or a combination of these effects. Let's go. Secondary prevention studies have shown that administration of long-chain n-3 PUFA to patients already suffering from myocardial infarction (MI) has shown significant benefits, which is EP- B-1152755 is claimed and claimed. n-3 PUFA is particularly powerful in reducing sudden death (see EP-B-1152755), which is an effect that occurs even without a significant reduction in lipids. This effect has been speculated to be due to the antithrombotic and antiarrhythmic action of n-3 PUFA. On the other hand, we have thought that n-3 PUFA contributes to the stabilization of atheroma plaques by its action against inflammation. Conversely, n-6 PUFA linolenic acid is found in vegetable oils such as sunflower oil, but can promote inflammation. In this case, increased intake of linolenic acid leads to instability of plaque. There are indications that it may be.

To our knowledge, no studies have reported the effect of n-6 or n-3 PUFA on plaque stability. Rapp et al. (See Arterioscler. Thromb. 1991, Vol. 2, pages 903-911) patients who are to undergo endarterectomy have received very high doses of fish oil (daily Studies were conducted at a rate of 48 to 64 grams per liter, but they were n-3 PUFA, eicosapentaenoic acid (EPA; 20: 5n-3), and docosahexaenoic acid (DHA; 22: The level of 6n-3) was found to be very high compared to in plaques removed from control patients. However, Rapp et al. Do not give details regarding the structure of the plaque.
European patent EP-B-1152755 Rapp et al. (Arterioscler. Thromb. 1991, Vol. 2, pages 903-911) Am. J. et al. Clin. Nutr. 2001, 73, 539-48 Story et al. Circulation, 1995, 92, 1355-74 Arterioscler. Thromb. Vasc. Biol. 2000, 20, pp. 1262-75

  In particular, we found that patients who were given fish oil orally tended to have more plaques covered by a better-shaped fibrous covering rather than a thin inflamed covering, He also discovered that macrophage infiltration into the plaque was not so severe. As is well known, features that make plaques susceptible to rupture atherosclerosis include that they have a thin fibrous covering and an increased number of inflammatory cells such as macrophages. The presence of macrophages in the carotid plaque is also known to be associated with an increased incidence of brain disease events, such as strokes and transient ischemic attacks. Therefore, oral administration of fish oil provides an effective and safe method to help prevent brain disease events, particularly stroke, in patients with arterial atherosclerosis symptoms of arteries supplying blood to the brain. is there.

  Accordingly, the present invention uses EPA and / or DHA in formulating oral medications to prevent damage to the brain of a patient who exhibits signs of atherosclerosis in the arteries that supply blood to the brain. Is directed to.

  In a preferred embodiment of the present invention, the oral medication is for preventing patients exhibiting symptoms of cerebrovascular disorders, transient cataracts and / or transient cerebral ischemic attacks from causing a stroke. .

  Here we describe in detail the experimental studies that led to the present invention.

[Patients and methods]
[Research Plan]
Two patients who have undergone carotid endarterectomy, in other words, who have advanced symptoms of atherosclerosis in the arteries that supply blood to the brain, who have agreed to participate in the experiment, A double-blind approach was to receive one of three types of oil provided in random capsules. The control oil is a mixture of palm oil and soybean oil in a ratio of 80:20, the fatty acid composition of which is matched to the average dietary fatty acid composition of an English adult. The other oils were sunflower oil and fish oil containing EPA / DHA. The fatty acid composition of the oil is listed in Table 1. Each capsule contained 1 gram of oil and 1 milligram of (α-tocopherol. Patients consumed 6 capsules a day until surgery, and 3 It was recommended to consume 2 capsules per meal, so the amount of long chain n-3 PUFA provided was 1.4 grams per day. Was 3.6 grams per day, which represents a 40 percent increase in intake of this fatty acid.

  Table 1. Fatty acid composition of capsules used in the study.

  The specimen size was determined based on the fatty acid composition of plaque phospholipid (PL) described in Rapp et al. (Arteriscler. Thromb. 1991, 11, 903-11). The DHA dose was calculated taking into account that it is 10 to 15 times lower. Based on this, a sample as large as 50 was calculated to be necessary to detect a 1-fold increase in EPA in plaque PL at P <0.05. In anticipation of a dropout ratio of up to 20 percent, the study recruited 188 patients.

  When participating in the study, fasting venous blood specimens were incorporated into EDTA. The patients then began consuming the capsules in the manner described above and continued to weigh and keep the diary for 7 days. During the study period, patients were advised to continue medication and not change their diet. Regular contact with the patients encouraged compliance monitoring and was assessed by counting the number of returned capsules and measuring the fatty acid composition of the LDL lipid fraction. Patients who reported that they were unable to comply (n = 5) withdrew from the study. Measurement of the number of capsules returned suggested that compliance was greater than 85 percent in any treatment group. Furthermore, the proportion of EPA in each plasma LDL lipid fraction increased by more than 0.5 grams in 100 grams of total fatty acids in over 90 percent of patients in the group receiving fish oil. These findings indicate that at least 85 to 90 percent compliance was achieved in each treatment group.

  Surgical excision of the carotid plaque was performed generally 7 to 190 days after the patient participated in the study. Patients who underwent surgery within 7 days were excluded from the study. The fasting venous blood sample was taken again the morning before surgery. During the surgery, carotid plaque was collected and cleaned. It was then cut into 2 mm wide sections starting from the common carotid artery. The cut surface for biochemical analysis was frozen in liquid nitrogen. The cut surface for histological analysis was fixed in formaldehyde and then embedded in paraffin wax. Sections for immunohistochemical analysis were frozen in an optimal cutting temperature embedding medium (Agar Scientific, Stansted, UK) and stored at -70 degrees.

[Analysis of habitual nutrition]
An ingested food diary was analyzed to find habitual nutrition intake using a modification of FOODBASE (Brain Chemistry Laboratory, London, UK), which was effective in determining fatty acid intake.

[Plasma lipids and lipoproteins]
Total concentrations of cholesterol and triacylglycerol (TAG) in plasma were determined using a commercially available colorimetric assay (Sigma Chemical Co., Poole, UK). Low density lipoprotein (LDL) is a solution of 1.7 milliliters of plasma (adjusted to a concentration of 1.24 grams / milliliter by adding solid potassium bromide) to 3.3 milliliters of phosphate buffered saline. The bottom layer was formed and then centrifuged in a Beckman TLA-100.4 rotor of a Beckman Optima ultracentrifuge at 15 degrees Celsius for 2 hours at a speed of 100,000 rpm in a sealed tube. Adjusted from plasma over step density gradient. Purified LDL was frozen at -70 degrees for fatty acid composition analysis.

[Fatty acid composition analysis]
The fatty acid composition of the frozen fragment closest to the LDL PL, cholesteryl ester (CE), and TAG fractions, and carotid plaque branching was determined. Total lipids were extracted, lipid fractions were separated by thin layer chromatography, and the fatty acid composition of each fraction was described by Thies et al. (Am. J. Clin. Nutr. 2001, 73, 539-48). Page) determined by gas chromatography.

[Carotid plaque morphology]
Fragments embedded in paraffin were stained with hematoxylone and eosin. The fragment closest to the bifurcation is described in guidelines published by the American Heart Association (Stary et al., Circulation, 1995, 92, 1355-74) and a modification of this system proposed by Virmani et al. (Arterioscler.Thromb .Vasc.Biol.2000, 20, pp. 1262-75). The fragments were examined by a cardiovascular pathologist (PJG) in random order without providing information about the patient. The American Heart Association (AHA) classification includes the following six classes or types: Type I (early disability); Type II (early disability or steatosis); Type III (intermediate disability or preatheroma); Type Includes IV (atheroma or atherosclerotic plaque); type V (fibrous atheroma or fibrosis); type VI (disorder with superficial abnormalities and / or bleeding and / or thrombus deposition). The revision of this classification proposed by Virmani et al. Is a series of grades with a description showing an increase in severity as follows: pathologic intima (increased smooth muscle cells in the matrix With area of accumulation, but no necrosis or thrombus); fibrous atheroma (with a well-shaped necrotic nucleus overlaid by fibers; no thrombus); thin fibrous covering Atheromas (thin fibrous coatings invaded by macrophages and lymphocytes, muscle cells are rarely smooth, necrotic nuclei are underneath, no thrombus); sputum (luminal thrombosis); plaque rupture (ruptured fibrosis) Atheroma, with luminal thrombus leading to necrotic nuclei); including lime nodules and fibrotic calcified plaques (rash calcification)

[Immunohistochemistry of carotid plaque]
The area of the plaque closest to the bifurcation was used for immunohistochemistry. The area consists of the presence of macrophages (identified by the presence of CD68 on the surface) and T lymphocytes (identified by the presence of CD3 on the surface), and two types of adhesion molecules: conduit cell adhesion molecules— Stained to detect 1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1), they are associated with movement of immune cells into plaques. The cryopreserved area of the frozen plaque was placed on a microscope slide coated with an organosilane compound. Endogenous peroxidase activity is blocked and then the zones are sequentially diluted with different dilutions of anti-human antibodies, namely biotinylated goat anti-mouse (pig anti-goat for staining VCAM-1) immunoglobulin G (DAKO, Ely, UK) and streptavidin-horseradish peroxidase (DAKO, Ely, UK). Finally, peroxidase activity was visualized using hydrogen peroxide as a substrate and 3-amino-9-ethylcarbazole (Sigma Chemical Co., Poole, UK) as a chromogen. Stained areas were fixed using formalin, counterstained using Harris hematoxylin, and observed at 10X magnification using a microscope. Primary antibodies used were mouse anti-human antibody CD3 (Leu 4, Becton Dickinson, Oxford, UK), mouse anti-human antibody CD68 (KP1, DAKO, Ely, UK), mouse anti-human antibody ICAM-1 (R (D Systems, Oxford, UK) and goat anti-human antibody VCAM-1 (R (D Systems, Oxford, UK). Staining was 0 (no staining), 1 (moderate staining), or 2 (dark staining). ).

[Statistical analysis]
Data are shown only for patients who completed the study (n = 57 in the reference group, n = 52 in the sunflower oil group, and n = 53 in the fish oil group). Age, body mass index (BMI), blood lipid level at the start of the study, medical history, drug use and habitual nutrition among different treatment groups using single factor analysis of variance (ANOVA) Compared. The effect of treatment on blood lipid concentration and fatty acid composition of LDL lipid fraction was determined as a change from baseline values. Changes from baseline between different treatment groups were compared by single factor covariance analysis (ANCOVA) using baseline values and duration of treatment as covariates, where the effect of treatment was Student t-tests were used to identify differences between groups when significant. In some cases, post-treatment values are compared to baseline values within the same treatment group by paired student t-test, and single-factor ANOVA (post-event student t-test) ANOVA) was used to compare post treatment values between treatment groups. Among different treatment groups, the fatty acid composition of carotid plaque lipid fraction was compared using single factor covariance analysis (ANCOVA) using the duration of treatment with oil as a covariate, with the effect of treatment Student t-test was used to identify differences between groups. Staining intensity (immunohistochemistry) and plaque morphology distribution were compared between treatment groups using the chi-square test. The average grade of each distribution within each treatment group was determined and these were compared by the Yonky Therapstra test. The correlation was determined as the Spellman correlation coefficient (ρ). All analyzes were performed using SPSS version 11 (SPSS, Chicago, Illinois, USA), and in all cases a value of P <0.05 was considered to indicate statistical significance.

[result]
[Patient characteristics]
Eighteen patients withdrew from the study, 13 for clinical reasons and 5 failed to follow the study procedure. An additional 8 patients were removed from the study because they underwent surgery within 7 days of the start of the study.

  The characteristics of the patients who participated in this study are shown in Table 2. Gender ratio, age, BMI, fasting plasma TAG and cholesterol levels, intake of individual macronutrients and micronutrients, including energy and individual fatty acids, number of smokers / former smokers among treatment groups at study registration There were no significant differences in terms of ratio, degree of stenosis of the diseased carotid artery, history and use of drugs (all Ps are at least greater than 0.1438. Single factor analysis of variance (ANOVA) )) (See Table 2). Each group received supplemental oil for the same period (see Table 2). The sunflower oil group of patients took an extra 3.6 grams of linolenic acid per day, but the consumption of this fatty acid increased by about 40 percent. Patients in the fish oil group consumed an extra 1.4 grams of long chain n-3 PUFA per day, but EPA intake increased approximately 10-fold and DHA intake increased approximately 4-fold.

  Table 2. Patient characteristics at study registration.

Age, BMI, blood lipid and nutrient intake data are mean values (standard deviations are shown (age and BMI ranges are shown in parentheses).
Data on stenosis are median and 25th and 75th percentile values shown in parentheses.
Data on the length of the oil treatment period are the median and the 10th and 90th percentile values shown in parentheses.
(Note 1) This data is calculated based on a diary that records the weight of food eaten over 7 days.
(Note 2) Most former smokers quit smoking more than three years before the registration.
(Note 3) This refers to a stroke.
(Note 4) Temporary, partial or complete blindness.

[Plasma lipid concentration]
There was no significant treatment effect on plasma cholesterol levels (data not shown). However, there was a significant effect on plasma TAG concentration (P = 0.0184; single factor covariance analysis (ANCOVA)). Fish oil administration resulted in a significant decrease in plasma TAG concentration (−0.48 ± 1.06 mmol / L), but this value did not change significantly in other groups. Therefore, TAG concentrations in plasma were significantly lower after treatment with fish oil (1.2 ± 0.8 mmol / L) compared to baseline values (P = 0.0032; paired student t-test). ). Furthermore, the plasma TAG concentration in the fish oil group at the end of treatment was significantly lower than that in the other two groups (P = 0.0294 for the reference group and P = 0.0347 for the sunflower oil group). Single factor analysis of variance (ANOVA)). There was a significant first-order correlation between the length of the treatment period using fish oil and the change in plasma TAG concentration ((= −0.44, P = 0.118).

[Fatty acid composition of LDL lipid fraction]
There was a significant therapeutic effect on the ratio of several fatty acids in LDL PL, CE and TAG. After fish oil treatment, the ratio of EPA and DHA increased in all three LDL lipid fractions (Table 3). These ratios are significantly different from baseline (P is less than 0.0001 for EPA in each fraction, P is 0.0031 for DHA in CE, P for DHA in PL and TAG. Was less than 0.0001; paired student t-test) and was also significantly different from the proportions in the other two groups after treatment (at least P ≦ 0.0003; single factor analysis of variance (ANOVA)). The increase in the ratio of long-chain n-3 PUFAs in LDL PL in the fish oil group is due to the ratio of linolenic acid, di-homo-γ-(-linolenic acid (20: 3n-6) and arachidonic acid (20: 4n-6). The increase in the ratio of long chain n-3 PUFA in LDL CE and TAG in the fish oil group was accompanied by a significant decrease in the respective ratios of linolenic acid and oleic acid (18: 1n-9). Treatment with sunflower oil resulted in an increased proportion of linolenic acid in LDL CE, which was mainly at the expense of oleic acid, and these findings are summarized in Table 3.

  Table 3. Effect of oil diet on fatty acid composition of LDL lipid fraction.

[Fatty acid composition of carotid plaque]
There was a significant therapeutic effect on the ratio of EPA and DHA in each of the plaque lipid fractions and the ratio of linolenic acid in plaque PL. The EPA ratio in PL, CE and TAG contained in carotid plaques of patients in the fish oil group was calculated as P <0.0001, 0.0053, and 0.0007 for the patients in the reference group (PL, CE, and TAG, respectively) Single factor covariance analysis (ANCOVA)), and sunflower oil group patients (P <0.0001, 0.0278 and 0.0024 for PL, CE and TAG respectively); single factor covariance analysis (ANCOVA) It was higher than that of)). There was a large positive first-order relationship between the EPA ratio in plaque PL and the length of treatment with fish oil ((= 0.41, P = 0.0051). Included in carotid plaques of patients in fish oil group The ratio of DHA in CE and TAG was higher than that of patients in the reference group (P was 0.0042 and 0.0241 for CE and TAG, respectively; single factor covariance analysis (ANCOVA)). The DHA ratio in PL and CE contained in the carotid plaque of the fish oil group patients is that of the sunflower oil group patients (P is 0.0100 and 0.0278 for PL and CE respectively; single factor covariance analysis ( Compared to ANCOVA)), the PL contained in the plaques of patients in the fish oil group includes the other two groups. A lower proportion of linolenic acid was found compared to this (P for the reference group was 0.0118; P for the sunflower oil group was 0.0015; single factor covariance analysis (ANCOVA)). There was no significant change in fatty acid composition in the plaque lipid fraction between the group and the sunflower oil group, and these findings are summarized in Table 4.
Table 4 Fatty acid composition in carotid plaque lipid fractions of different oil treatment groups.

Data are mean ± standard deviation.
Values in a row indicated by different superscripts are significantly different from each other (single factor covariance analysis (ANCOVA) using length of treatment period as a covariate).

[Morphological physiology classification of carotid plaque]
The distribution of disability forms determined using the AHA classification is significantly different between patients taking fish oil and patients taking reference oil or sunflower oil (P vs. reference oil group). 0.0234, P for sunflower oil group is 0.0107; Chi-square test). This is thought to be due to the greater proportion of type IV disorders (“atheroma”) and the lower proportion of type V (“fibroatheroma and fibrotic disorders”) in the fish oil group. We compared the distribution of disability types among patients who were treated shorter or longer than the intermediate treatment period in each group. For plaques of patients who received treatment shorter than the intermediate treatment period, the distribution was not significantly different between treatment groups. However, the disability distribution of patients who have been treated with fish oil for longer than the intermediate treatment period is found in both the reference oil group and the sunflower oil group (P is 0.0111 and 0.0432, respectively, chi-square test). Significantly different from that observed. Over all patients, EPA and DHA content was highest for type IV plaques and lowest for type VI plaques.

  The disability type distribution determined using the modified AHA classification was significantly different between patients taking fish oil and those taking reference oil or sunflower oil (for reference oil groups, P is 0.0344, for sunflower oil group, P is 0.0313; Chi-square test). The difference is that in the fish oil group, there is a greater proportion of neatly shaped fibrous covering disorders, the absence of thrombus (fibrous covering atheroma), and thin, inflamed fibrous This is probably due to the lower proportion of wrapping disorders (thin fibrotic wrapping atheroma).

  There was no significant difference in the distribution of plaque disorder types between the reference oil group and the sunflower oil group. These findings are summarized in Tables 5 and 6.

  Table 5 Carotid plaque morphology in different oil treatment groups.

(Note 1) The disability type distribution is significantly different between the fish oil group and the reference oil group (P is 0.0234), and is also significantly different between the fish oil and sunflower oil groups (P is 0.0107) ( Chi-square test).
(Note 2) The disability type distribution is significantly different between the fish oil group and the reference oil group (P is 0.0344), and is also significantly different between the fish oil group and the sunflower oil group (P is 0.0313). (Chi-square test).

  Table 6 n-3 polyunsaturated fatty acid content and macrophage (anti-CD68) staining intensity in carotid plaque lipids according to AHA classification.

AHA classification values indicated by different superscripts are significantly different from each other (single factor covariance analysis (ANOVA)).
* Indicates that anti-CD68 staining intensity 2 is different from staining intensity 1 (Student's t-test with no correspondence).

[Lymphocytes and macrophages in carotid plaque]
There was no difference regarding the presence of ICAM-1 or VCAM-1 in plaques taken from patients in different treatment groups (data not shown). Similarly, there was no therapeutic effect on the presence of T lymphocytes in the plaque. In contrast, plaque fragments from patients who ingested fish oil are somewhat less likely to be stained with the macrophage marker anti-CD-68, and the distribution of staining points for this group is different compared to the other groups (relative to the reference oil). P <0.0001, P for sunflower oil was 0.0016; Chi-square test), and the average rank of staining intensity was significantly lower (P was 0.0246). The length of the treatment period with fish oil is significantly inversely related to the intensity of anti-CD-68 staining ((= -0.352, P = 0.0301). Intake of fish oil for a shorter period than the average treatment period. The plaques of the treated patients showed higher anti-CD68 staining intensity (staining intensity 47 than staining intensity 1 and 53% staining intensity 2) compared to those treated longer than the average treatment period (staining intensity). (Intensity 1 was 25 percent and staining intensity 2 was 75 percent.) However, there was no significant change in the distribution of these staining intensities (P was 0.0827; Chi-square test). We compared the distribution of anti-CD68 staining intensity in plaques taken from patients who were treated for shorter or longer periods than the mean treatment period.The distribution observed in the fish oil group was the reference oil group (mean treatment period). P <0.0001 and 0.0048) for each group of patients who received shorter or longer periods of treatment, and sunflower oil groups (shorter than average treatment period) Significantly different from that of both P <0.0109 and 0.0363) for each group of patients who received or for long periods of time (chi-square test). Permeated plaques (ie anti-CD68 staining intensity 2) contain very little EPA and DHA compared to mildly permeated plaques (ie anti-CD68 staining intensity 1) (Table 6).

  There was no significant difference between the reference oil group and the sunflower oil group in terms of anti-CD68 staining score or average rank distribution of staining intensity.

  The findings of T lymphocyte and macrophage staining are summarized in Table 7.

  Table 7. T lymphocyte and macrophage staining in carotid plaques of patients belonging to different oil treatment groups.

(Note 1) The distribution of the staining score is significantly different between the fish oil group and the reference oil group (P <0.0001) and between the fish oil group and the sunflower oil group (P = 0.016) ( Chi-square test).
(Note 2) The numbers in one column indicated by different superscripts are significantly different from each other (P = 0.0246; Yonky Therapstra test).

[Discussion]
In this study, adding sunflower oil to the patient's diet provided 3.6 grams of linolenic acid per day, but had a very limited impact on the measured results. This is probably due to the fact that these patients had already consumed a significant amount of linolenic acid in a habitual diet, but this intake is consistent with what has been reported for British adults. In our findings, at least for the duration of the study, subjects with advanced carotid atherosclerosis increased their linolenic acid intake to 40 (and the consumption of a standard amount of this fatty acid in carotid plaques. It was shown that it did not lead to an increase in linolenic acid uptake, nor a change in plaque stability.

  Adding fish oil to the diet significantly reduced plasma TAG levels. The degree of TAG reduction was consistent with that seen in other studies on fish oil additions and was related to the length of time the fish oil was added.

  Long chain n-3 PUFAs such as EPA and DHA are usually consumed in small quantities and are therefore found in relatively low proportions in plasma and tissue lipids. However, the increase in consumption of these fatty acids can be seen from their increased ratio in various blood and tissue lipid retention, as observed in this study on LDL lipid fractionation. A key finding of this study is that when small amounts of long-chain n-3 PUFA are consumed, they are readily incorporated into atherosclerotic plaque lipids. Incorporation of EPA into plaque lipids, especially PL, is first order over time. To the best of our knowledge, the only previous study that examined the effect of fish oil supplementation on fatty acid composition in atherosclerotic plaques was the above study by Rapp et al., Where it continued when large amounts of fish oil were consumed, It was stated that large amounts of EPA and DHA were incorporated into plaque lipids. However, Rapp et al. Did not examine the effects of EPA / DHA intake on plaque morphology. In the present invention, n-3 PUFA administered to the same extent as that used in research on breakthrough secondary prevention (see eg EP-A-1, 152, 755) is incorporated into plaque lipid retention. Prove that In addition, uptake of n-3 PUFA occurs within a relatively short time frame, even at the small doses that we have done. This suggests that atherosclerotic plaques are fairly dynamic and that some degree of lipid turnover can be seen even at a stage where atherosclerosis has advanced considerably.

  Immunohistochemical staining and plaque morphology measurements revealed that n-3 PUFA has great influence. In the fish oil group, there were more plaques than the other two groups with a cleanly shaped cover rather than a thin inflamed cover. Furthermore, plaques from patients treated with fish oil were not severely invaded by macrophages. The changes in plaque morphology observed as a result of the addition of fish oil in this study thus indicate that the more stable plaques, ie the risk of rupture, have become lower. These differences (i.e., less macrophage invasion and increased number of plaques covered with a cleanly shaped fibrous covering) contained more EPA and DHA in plaque lipids. This indicates that it is these n-3 PUFAs that determine plaque stability.

[Formulation and administration form of drug]
In the present invention, the active ingredient of the drug is EPA, DHA, or a mixture of EPA and DHA. These n-3 PUFA fatty acids may be present in the form of naturally occurring triglycerides and also take the form of pharmaceutically acceptable salts or derivatives, especially their ethyl esters or other alkyl esters. Sometimes.

  While other EPA and DHA ingredients may be commercially available in the future, the active ingredient is derived from fish oil in an appropriate manner. Methods for producing pharmaceutical grade oil from raw fish oil and methods for incorporating various concentrations of EPA and / or DHA into the product are known to those skilled in the art. Increase the EPA and / or DHA concentration in fish oil, for example in the form of one or two capsules daily, in order to increase the rate of n-3 PUFA intake and ensure patient compliance Is preferably provided. A composition containing 20 to 100 weight percent of a mixture of EPA and DHA is preferably used as an active ingredient, and a composition containing 70 weight percent or more of a mixture of EPA and DHA is more preferably used. Most preferably, a composition containing 70 to 90 weight percent EPA / DHA mixture is used. Although it is not believed that the ratio of EPA to DHA in the mixture is significant, it is usually preferred that the relative amount of EPA: DHA is 1: 2 to 2: 1, approximately 3: 2. It is more preferable.

  While it is preferred that the active ingredient of the drug contains a mixture of EPA and DHA, within the scope of the present invention, it is noted that each of these n-3 PUFA acids is used alone. Methods for formulating essentially pure EPA or DHA are known and are described in the literature.

  The drug of the present invention is for oral administration. Other oral dosage forms may be used as needed, for example powders obtained by microencapsulation, but oral dosage forms are suitably capsules with a hard or soft shell.

  In addition to the EPA and DHA active ingredients defined as active ingredients, the medicament may include one or more carriers that can be used as drugs known to those skilled in the art. The composition may also contain fillers, stabilizers, bulking agents, binders, humidifiers, surfactants, lubricants and the like, as shown in the art of determining chemical composition.

  In addition, for example, hydroxytoluene, butyrate, quinone, tocopherol, ascorbic acid and other antioxidants, preservatives, colorants, fragrances, artificial sweeteners and other chemical ingredients can also be used. Antioxidants are a particularly preferred optional ingredient for drugs.

[Examples of oral drug formulation]
[Soft gelatin capsule containing 1 gram per capsule]
composition:
EPA ethyl ester 525 mg / capsule DHA ethyl ester 315 mg / capsule d- (tocopherol 4 immune unit / capsule gelatin 246 mg / capsule glycerol 118 mg / capsule The active ingredients and excipients are weighed and using a high speed stirrer The mixture was then subjected to a colloid mill and degassed in a stainless steel container ready for capsule filling.The mixture was placed into a soft gelatin capsule of size 20 oval (average weight). 1.4 grams), and filled using a conventional encapsulation machine.

  The agents of the present invention can be administered at any suitable dose to patients who exhibit symptoms of atherosclerosis in the arteries that send blood to the brain, but at a very high level of n-3 PUFA. This is because it is essentially non-toxic. In the test administration described above, dose management was done in such a way that a total of about 1.4 grams of EPA and DHA was provided per day, which was effective in improving plaque stability. It was shown that Generally, the dosage will be 0.5 to 5.0 grams of EPA and / or DHA per day, preferably 1.0 to 3.0 grams per day.

  The agents of the present invention provide additional, possibly fatal, seizures to patients presenting with symptoms of atherosclerosis in the arteries supplying blood to the brain, such as strokes or transient ischemic attacks. Suitably administered for the purpose of reducing the risk. To this end, the drug can also be used in combination with other therapeutic agents, such as aspirin and warfarin, known to reduce the risk of secondary brain disease events occurring in such patients. .

Claims (13)

In formulating oral medications to prevent brain damage in patients with atherosclerosis symptoms in arteries that supply blood to the brain,
Use of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or a mixture of EPA and DHA, or a mixture of salts or derivatives that can be used as drugs. Use according to claim 1,
Use characterized in that a mixture of EPA and DHA is used. Use according to claim 2,
Use wherein the ratio of EPA to DHA in the mixture is in the range of 1: 2 to 2: 1. Use according to claim 3,
Use characterized in that the ratio is approximately 3: 2. Use according to any of claims 1 to 4,
Use wherein both or any of said EPA and DHA are present as ethyl esters. Use according to any of claims 2 to 5,
Use wherein the medicament contains the mixture as an active ingredient in the range of 20 percent to 100 percent. Use according to claim 6,
Use wherein the medicament contains more than 70 weight percent of the mixture as an active ingredient. Use according to claim 7,
Use wherein the medicament contains 70 to 90 weight percent of the mixture as an active ingredient. Use according to any of claims 1 to 8,
Use, characterized in that the medicament also contains an antioxidant for the EPA and / or DHA. In formulating oral medications to prevent stroke in patients with cerebrovascular disorders, transient cataracts and / or transient cerebral ischemic attacks,
Use of EPA, DHA, or a mixture of EPA and DHA, or a mixture of salts or derivatives thereof that can be used as drugs. Use according to any of claims 1 to 10,
Use wherein the medicament is administered at a dosage of 0.5 to 5.0 grams per day. Use according to claim 11, comprising:
Use wherein the medicament is administered at a dosage of 1.0 to 3.0 grams per day. Use according to any of claims 1 to 12,
Use, characterized in that the oral medicine takes the form of a capsule.