Agent of hindu lotos seed (nelumbo nucifera) extract possessing anxiolytic and antidepressive action. RU patent 2497538.

IPC classes for russian patent Agent of hindu lotos seed (nelumbo nucifera) extract possessing anxiolytic and antidepressive action. RU patent 2497538. (RU 2497538):

A61P25/24 - Antidepressants

A61P25/22 - Anxiolytics

A61K36/28 - PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J0003000000; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L)

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FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely preparing an agent of hindu lotus seed extract (Nelumbo nucifera) possessing anxiolytic and antidepressive action. The agent possessing anxiolytic and antidepressive action prepared by extraction of hindu lotus seed extract (Nelumbo nucifera) in 50% ethanol in a Soxhlet extraction apparatus in the specific proportions.

EFFECT: agent possess the effective anxiolytic and antidepressive action.

3 tbl, 2 ex

The invention relates to the pharmaceutical industry, namely the creation of funds on the basis of the extract of the seeds Lotus (Nelumbo nucifera), which has anxiolytic and antidepressant effect.

According to who, anxious-depressive state, being widespread manifestation of stress-related diseases of the Central nervous system, there are more than 120 million people worldwide. According to some researchers, depending on the applicable criteria, anxiety as a pathological condition occurs in 5-25%, and mixed anxiety-depressive disorders are found in 13% of the population [3]. Increase the alarm and anxiety-depressive conditions associated with increased stress effects in modern society threatens not only the mental health of the population, but also provokes the development of psychosomatic diseases. It is well known that almost always mental equivalents of this mental disorder combined with somatic complaints, often with impaired function of the cardiovascular, immune and endocrine systems. Therefore, the search for the ways to correct these pathological States is one of the most important tasks of modern pharmacology. Despite the fact that modern medicine has learned how to cope with the state of anxiety and depression, there are problems of therapeutic resistance to popular psychotropic drugs and the formation of serious complications pharmacotherapy of affective disorders. Agents of vegetable origin can play a significant role in solving these problems.

Currently has the following known anxiolytic and antidepressant funds of natural origin: «Despres» and «» (St. John's wort); «Hyper» (St. John's wort, , hawthorn, vitamin C, vitamin A, vitamin B6 , vitamin B12 , magnesium carbonate); «» (passion fruit, hawthorn, Valerian, vitamin B1 , vitamin B6 , vitamin B 12 ). Close to the offered invention is a tool that has anxiolytic and antidepressant effect of the dried crushed aboveground and underground parts of species of the genus (Alfredia) or water or water-ethanol extracts (RF patent №2354397). However, the identical combination of traits us not found in the patent and scientific and medical literature.

The present invention is the creation of a domestic funds based on plant with an anxiolytic and antidepressant effect, has a wide range therapeutic activity. First established that as a means of having an anxiolytic and antidepressant effect, you can use water-alcohol extract of the seeds Lotus (Nelumbo nucifera).

Nelumbo nucifera is used in folk medicine since ancient times. The main biologically active substances extract of the seeds of the Lotus is flavonoids (quercetin, , ), (, ), alkaloids (). The fruit of the Lotus are members of more than two hundred products the Chinese and Indian medicine. Seed extract Lotus has hepatoprotective and antioxidant action. Nelumbo nucifera is a diuretic, febrifuge, hypoglycemic activity. In the literature there are no data on the use of water-alcohol extract of the seeds Lotus as a means having anksioliticski action.

Way to make an extract of the seeds Lotus (Nelumbo nucifera), obtained by means of water-alcohol extraction by 50% ethyl alcohol at a ratio of 3:1 in the device ml Soxhlet extractor (according to the Protocol WHO CG-04) [4]:

500 g of seeds were dried, crushed to powder-like state with the size of particles of 0,2 mm and subjected to water-spirit extracted 50%ethyl alcohol in the device ml Soxhlet extractor, then distilled alcohol in a rotary evaporator, in the following ratio: wt.%

Lotus seeds (Nelumbo nucifera)

- 75

ethyl alcohol

- 25

Output of the active substance (total alkaloids) extract, around 7% of viscous substances.

Solution of the task of the present invention, the use of the extract of the seeds Lotus on the proposed appointment has been made possible thanks to the experimental determination of its new properties, namely the ability to normalize the function of the CNS by anxiolytic and antidepressant activity.

Example 1.

Was followed by chemical analysis of the obtained extract seeds Lotus . By high performance liquid chromatography studied the composition of alkaloids contained in the extract. According to our chemical analysis of the main active components Lotus seeds when used method of extraction were the following alkaloids (in percentage):

27,14

21,87

14,73

from 14.25

11,02

6,21

Procyanidin 4,78

Assessment anxiolytic activity extract of the seeds Lotus spent 92 white the mongrel of adult rats-female study the peculiarities of behavior in « test. Animals were kept in vivarium with free access to water and food.

-test is a «hybrid» several behavioral models that allow you to maintain a wide range of behavioral reactions of locomotion and research activity up to the markers of autonomic behavior. As a completely new behavioral model, -test is recommended in psychopharmacology for the screening of psychotropic effects of drugs [2]. About anxiolytic activity extract judged by the following indicators: the latent period of exit from the center; the time spent in the dark and light compartments -test; quantity and duration ; number of drops down; the number of slipping back legs; number of stops near the border between black and white compartment. Additional indicators of conditioned-reflex, emotional and biased activity, served the number of visited segments; counters; the number and duration of stops; number of research down; the amount directed to the side movements of the head at a stretch position of the body.

Solution extract of the seeds Lotus (0.5%), prepared on the physiological solution, administered to animals intragastric tube at the dose of 50 mg/kg of body weight. The animals were divided into three groups: I - intact control, II - animals receiving the intragastric physiological solution, III - animals receiving the intragastric solution extract of the seeds Lotus . The animals received solution extract of the seeds Lotus and saline solution every day at the same time, from 10 to 12 hours. Seed extract Lotus and physiological solution is administered to animals for 1, 2, 3, 4 and 6 weeks and accordingly subdivided animals on the 1-, 2-, 3-, 4 - and 5-th group. Each group of animals tested in test once within 30 minutes after the last intravenous injections.

The obtained experimental data were statistically processed using t criterion adjusted Б. Differences was considered significant at p≤0,05.

Our studies (table 1, 2) showed that the extract of seeds Lotus leads to a decrease in latency time exit from the center; reduction of the number of slippage paws; reduction of defecation, since the first week of intragastric administration of the extract of the seeds Lotus compared with intact animals. While in animals treated with saline is increasing the level of anxiety in response to the procedure intragastric administration (manipulations with animals, taking in hands), especially the first and second weeks of the introduction of physiological solution. This is manifested in the reduction of locomotor and exploratory activity, increase of time grooming and length of stops, and the appearance of falls and especially in the bright compartment.

Table 1

Dynamics of indicators of behavioral reactions in test with the introduction of Lotus extract of the seeds

1 week 2 week 3 week 4 week 6 week control

Latency output, s

3,9±0,56 *

2,7±0,47 *

4,0±0,60 *

18,9±11,48

24,5±18,83

11,8±3,05

The duration of stops, s, ON/ABOUT

61,2±27,45

34,5±9,81

33,67±and 11.38

11,5±of 7.64

62,0±18,94

20,1±10,23

Vertical activity

14,0±5,01

16,5±3,39

16,22±4,99#

10,3±5,66 ▲

12,75±4,46

24,3±4,73

20,6±5,12

26,5±5,83

14,3±3,95 #

40,0±1,17

28,4±7,29

Horizontal activity

0,3±0,15

0,22±0,22

0,1±0,1 0

0,8±0,36

0,3±0,21

0,3±0,15

0 0

0,6±0,44

Stop

1,9±0,70

2,4±0,65

1,7±0,48#

0,8±0,61 ▲ ▲

1,8±0,32

2,8±0,95

3,0±0,76

2,3±0,68

3,1±0,95

3,2±0,61

2,4±1,00

Looking down

3,3±0,99

5,3±1,18

5,4±1,95

3,6±1,38

5,2±1,33

6,9±1,75

4,5±1,44

4,7±1,22

5,1±1,17

4,4±1,40

6,4±1,55

Orientation

0,4±0,31 *▲

3,1±0,71 about

1,22±0,36

0,8±0,42 *

0,6±0,31 **

2,4±0,64

3,5±1,10

4,04-1,02

1,33±0,47

1,5±0,56

2,2±0,78

The time spent in the compartment, sec

64,9±18,76 **

139,3±29,64

117,2±24,41 #

69,6±35,93

117,8±28,52

160,3±the 28.38

134,7±31,06

127,9±28,75

147,4±32.76ˆ

152,7±40,03

120,2±29,32

Ectopic activity, grooming, sec

12,0±6,41

11,5±4,99

18,56±6,44

3,0±2,79

4,42±4,15

4,3±2,08

9,8±5,05

7,0±3,81

of 15.6 6,56

22,4±17,7

5,8±3,88

Defecation, the number of boles

0,3±0,21

1,4±0,99

0,44±0,34

0,3±0,21

0,25±0,18

0,6±0,27

0,6±0,34

0,1±0,11

1,3±0,49

0,1±0,10

Duration of stops, sec

21,0±9,9

51,3±7,89

60,6±24,14

34,0±24,65

58,7±24,12

41,8±12,38

31,8±9,08

32,1±13,93

58,0±21,97

87,4±29,32

26,1±11,47

Fall 0 0 0 0 0

0,1±0,10

0 0 0 0 0 0

Slippage paws

0,7±0,42

0,6±0,29 ▲

1,2±0,32

0,6±0,34

1,6±0,45

1,2±0,36

1,5±0,56

0,5±0,22

1,0±0,44

1,2±0,39

2,0±0,60

Table 2

Dynamics of indicators of behavioral reactions in test with the introduction of physiological solution

1 week 2 week 3 week 4 week 6 week control

Latency output, s

6,0±1,76

3,5±0,67 *

23,0±9,81

5,3±1,36

11,8±4,97

11,8±3,05

The duration of stops, s, ON/ABOUT

22,6±1,01

28,1±10,42

27,6±22,26

22,3±8,72

94,4±23,21 *

20,1±10,23

Vertical activity

22,0±of 10.52

22,3±4,41

4,4±3,97 **

9,5±3,28 *

11,9±5,73 *

24,3±4,73

22,2±make up 9.64

14,3±3,24

3,8±2,85 **

27,4±5,53

28,4±7,29

Horizontal activity

0,6±0,27

0,3±0,21

0,3±0,15

0,8±0,36

0,8±0,62

0,5±0,30

0,3±0,30

0,6±0,44

Stop

2,66±1,01

2,3±0,62

0,2±0,13 **

1,6±0,65

2,8±0,95

4,2±1,44

1,5±0,82

1,3±0,58

4,4±0,73

2,4±1,00

Looking down

2,4±0,87 **

7,6±1,76

2,9±0,95 *

4,3±0,97

7,7±1,32

6,9±1,75

4,9±2,74

3,6±0,73

0,7±0,37

6,9±1,38

6,4±1,55

Orientation

1,7±0,69

2,2±0,55

1,1±0,48

1,2±0,42

2,4±0,92

2,4±0,64

3,4±1,19

4,5±0,56

0,4±0,34

6,3±1,4

2,2±0,78

The time spent in the compartment, sec

of 116.7±37,47

to 156.2±30,55

38,4±27,36 **

23,2±7,32 **

115,9±26,85

160,3±the 28.38

108,1±42,48

116,8±32,83

21,33±21,33

215,7±26,64

120,2±29,32

Ectopic activity, grooming, sec

7,9±7,28

4,51±3,01

3,7±2,56

7,0±3,42

4,3±2,08

4,88stick 3,23

4,1±4,10

9,6±4,74

5,8±3,88

Defecation, the number of boles

0,7±0,37

0,5±0,27

1,6±1,16

0,3±0,28

1,4±0,75 *

0,6±0,27

0,2±0,16

0,6±0,49

0,7±0,42

0,1±0,10

27,1±16,23

41,4±13,32

29,0±26,85

29,2±13,24

41,8±12,38

Duration of stops, sec

22,0±7,76

31,8±19,44

71,0±36,11

47,8±12,47

26,1±11,47

Fall 0

0,2±0,13

0 0

0,2±0,13

0,1±0,10

0 0 0 0 0

Slippage paws

1,1±0,54

2,3±0,58 #

1,1±0,46

1,4±0,39

3,3±0,75 *

1,2±0,36

1,5±0,65

0,8±0,33

2,7±0,68

2,0±0,60

* as compared to the control; s - between black and white compartment; compared with the previous group; # - between Lotus and physiologic solution

While seed extract Lotus , shows anxiolytic action, the behavioral responses of change not only about the reactions of animals receiving saline solution, but relatively intact rats, indicating the effectiveness of the extract of the seeds Lotus as anxiolytic funds.

Example 2.

Assessment of antidepressant action of the extract of the seeds Lotus held on 24 white mongrel Mature rats female in the test of «» (swimming test «despair»). Animals were kept in vivarium with free access to water and food.

Test », the recommended when conducting psychopharmacological research to identify the antidepressant properties of pharmacological substances [1]. Installation is a glass cylinder (d=20 cm and a height of 40 cm, 2/3 full of water (t=25±1 C). The animal is placed in the cylinder and the following parameters are registered: active and passive (drift+full immobility) navigation, the latent period (LP) until the signs of the first and the first movement, the duration of . Follow-up time: 3 minutes

Solution extract of the seeds Lotus (0.5%), prepared on the physiological solution, administered to animals intragastric tube at the dose of 100 mg/kg body weight. The animals were divided into three groups: I - intact control, II - animals receiving the intragastric physiological solution, III - animals receiving the intragastric solution extract of the seeds Lotus . The animals received solution extract of the seeds Lotus and saline solution every day at the same time, from 10 to 12 hours. The duration of administration of the extract of the seeds Lotus and physiological solution was 8 weeks. Each group of animals tested in the test of «» once within 30 minutes after the last intravenous injections.

The obtained experimental data were statistically processed using t criterion adjusted Б. Differences was considered significant at p≤0,05.

Performed by us studies (Table. 3) showed that the extract of the seeds Lotus leads to a significant decrease latent period before the first movement and the first , as well as changes in the ratio of active/passive passage» by increasing the active component in comparison with intact animals, indicating that the increase on a background of the introduction of the extract of the seeds Lotus physical endurance in «» the aquatic environment and to show them expressed antidepressant properties.

Table 3

The study of the antidepressive activity of extract of the seeds Lotus in the test of «»

Group

PL before 1st movement, Pitch m, with

PL until the 1st , Pitch m, with

Time active swimming, Pitch m, with

Time passive swimming, Pitch m, with

Mobility, Pitch m, with

Control 1

5,2±0,6

59,2±6,2

120,3±3,5

41,3±2,7

16,7 ą 1.3

Control 2 (Phys. R-R)

16,4 of + 2,1Δ

60,5±5,9

117,4±4,3

60,0 of - / + 6,4Δ

18,8±1,2

Seeds Nelumbo nucifera (100 mg/kg)

7,8 ą1.5*

87,8±9,4*

131,2±4,3*

27,4±3,5*

21.4 ħ 1,9

Legend: Δ and * - p<0.05 - reliability of differences compared with controls 1 and 2, respectively (student's t-criterion adjusted Б)

2. A.V. -test a new behavioral model anxiety / AV , P. // Neuroscience. - 2005. - №1. - P.17-23.

3. Santana L. A review of studies concerning treatment adherence of patients with anxiety disorders / L. Santana, L.F. Fontenelle // Patient Prefer Adherence. - 2011. - №5. - P.427-439.

4. WHO: protocol CG-04. Preparation of alcoholic extract for bioassay and phytochemical studies (APJF/IP, 1001 (A). Geneva, World Health Organization, 1983.

Tool that has anxiolytic and antidepressant effect, on the basis of extract of the seeds Lotus (Nelumbo nucifera), which is characterized by the fact that it is obtained by means of water-alcohol extraction 50%ethyl alcohol in the device ml Soxhlet extractor with the following component ratio, mass%:

Lotus seeds (Nelumbo nucifera)

ethyl alcohol

25,

output active extract of about 7% of viscous substances.