Bacopa monnieri complexes with phosphatidylserine and phosphoryl ethers and their use to enhance cognitive functions and memory

Abstract

Bacopa monnieri complexes with phosphatidylserine and phosphoryl ethers and their use to enhance cognitive functions and memory. The invention relates to extracts of Bacopa monnieri complexed with phosphatidylserine in the presence of phosphoryl ether compounds, such as phosphorylserine and / or phosphorylcholine. The complex, which optionally contains other phospholipids and other active ingredients, can be included in pharmaceutical compositions or food supplements and improves cognitive function, memory speed, and memory quality, while reducing mental fatigue. The invention is aimed at individuals requiring cognitive enhancement, particularly middle-aged and elderly people, as well as school-aged children.

Description

Bacopa monnieri complexes with phosphatidylserine and phosphoryl ethers and their use to enhance cognitive functions and memory Background of the invention With increased life expectancy worldwide due to improved healthcare, people now live longer than at any other time in human history. As a consequence, new challenges arise with aging. Aging is frequently associated with cognitive decline, even in otherwise healthy individuals, creating difficulties in social interaction. The current increase in the aging population represents an emerging public health issue related to disability, and there is a great need for new, safe drugs or plant extracts that enhance cognitive function in older adults to improve the quality of life for those with cognitive impairment. Among the numerous phytopharmaceuticals and their combinations used for this purpose, a special extract of Bacopa monnieri, commonly known as water hyssop, has shown some effects on memory impairment. Bacopa monnieri (L.) Pennel has been used for centuries in Ayurvedic medicine, either alone or in combination with other medicinal herbs, as a memory and learning enhancer, sedative, anxiolytic, and antiepileptic. Bacopa monnieri, a plant belonging to the Scrophulariaceae family and commonly known as Brahmi, grows naturally throughout Southeast Asia, where it has a long history in traditional medicine for its memory-enhancing and anxiety-reducing properties. The medicinal herb is currently marketed worldwide as a cognitive enhancer and blood sugar regulator. The plant contains a large number of active compounds, including alkaloids and saponins with different main structures, the principal active compounds being the steroidal saponins bacosides; the plant also contains cucurbitacins, iridoids, and tyrosol derivatives. Animal studies of whole-plant or alcoholic extracts of Bacopa monnieri have reported cognitive-enhancing effects, including improved motor learning and enhanced memory acquisition, consolidation, and retention in rats. These memory-enhancing effects have been attributed to saponins (bacosides, bacopasides, or bacopasaponins). Bacopasaponin constituents have been shown to facilitate mental retention in the avoidance response in rats and to reverse the amnesic effects of neurotoxin, scopolamine, phenytoin, electroconvulsive therapy, and immobilization stress. Hypothesized mechanisms of action include anti-acetylcholinesterase and anti-butyrylcholinesterase activities, upregulation, modulation of gamma-aminobutyric acid, antioxidant effects, protein synthesis in the brain, 5-HT agonism, and modulation of stress hormones in the brain. Bacopa extracts have also been shown to mitigate the neurotoxic effects of certain metals and nicotine, and to reduce p-amyloid levels in the brains of a transgenic double-cell rapid amyloid deposition (PSAPP) mouse model, suggesting mechanisms of action relevant to Alzheimer's disease. Some Bacopa monnieri extracts have also been used in the treatment of Alzheimer's disease in combination with specific drugs. The ethanolic extract increased cerebral blood flow, and other studies have described its antioxidant potential, which helps mitigate the adverse effects of oxidative stress on neurons. Regarding human trials, numerous clinical trials have been conducted over the past 20 years, many of them double-blind, but the results regarding efficacy are inconsistent. In the various studies, the complete characterization of the extract used, in terms of quality and saponin or active ingredient content, is never reported, and this may explain the variability in the results. In a recent meta-analysis, selected studies suggested that plant extracts could show some improvement in healthy subjects, as determined in verbal auditory learning tasks, reverse number recall, inspection time tasks, and working memory, although this improvement was not statistically significant. Bacopa extract showed moderate improvements in subjects with memory loss, although only a few of the neuropsychological tests showed statistical significance.Although a few studies show positive results for some parameters, there are only a limited number of studies to establish systematic protective and memory-enhancing effects of Bacopa extracts. Examples of clinical trials that apparently report positive effects but were criticized in meta-analysis have been reported. Among them, alcoholic extracts of Bacopa monnieri have shown memory-enhancing effects in three double-blind, randomized, placebo-controlled studies. One trial in 46 healthy volunteers aged 18–60 years assessed their performance on a series of cognitive tests after 5 weeks and after 12 weeks of 300 mg of B. monnieri extract daily. The researchers reported significant improvements in measures of the Rey Auditory Verbal Learning Test (AVLT) and in state anxiety. Another 3-month study in 76 healthy adults, aged 40–65 years, showed an effect of Bacopa on the retention of new information in delayed recall of word pairs.A third trial in people over 55 with age-related memory decline, again with 3 months of treatment, showed further improvements at 12 weeks on subsets of the Wechsler Memory Scale, with no loss of cognitive gains 4 weeks after the end of active treatment. Conversely, in a single-dose study of Bacopa extract in which neuropsychological tests were performed 2 hours after administration, no difference was found between groups. Two uncontrolled trials have reported memory- and learning-enhancing effects of Bacopa with long-term dosing in children and in patients with anxiety neurosis. Many researchers have conducted chemical characterizations of the plant. Currently, no data are available on the pharmacokinetics of Bacopa or its saponins. Phase I clinical studies confirmed the safety of bacosides in healthy male volunteers with both single and chronic dosing administered over a period of 4 and 6 weeks, respectively, at doses up to 450 mg of dry aqueous extract. An aqueous extract of Bacopa administered orally at up to a dose of 5 g / kg showed no toxicity in rats. The LD50 of an orally administered alcoholic extract was 17 g / kg. In the cited trials, total adverse events (AEs) were similar in the placebo and treatment groups. Combinations of Bacopa extract with phospholipids have been reported in the literature, with conflicting results regarding potential improvements of the combination compared to the extract alone. Unfortunately, the previously reported variability in results obtained using this technique is common for drugs of natural origin containing a large number of potentially effective substances, in varying amounts depending on the plant origin, the preparation of the extracts, and the formulation method; this last aspect is of considerable importance in ensuring the actual bioavailability of the administered active ingredients. Furthermore, determining a therapeutically useful effect on memory enhancement remains a complex task, requiring satisfactory results in a multitude of tests relevant to quantitative and / or qualitative functions, and so far only a few drugs have met this requirement. Therefore, there remains a need for new medications that have a strong and consistent effect on memory deficits and are effective across a broad spectrum of cognitive abilities. There is also a need to improve the effects of Bacopa in humans, which have thus far proven to be moderate in severity and inconsistent in occurrence, with the aim of ensuring the safe and highly effective use of this active ingredient according to modern medical standards. Summary of the invention It has now been found that an extract of Bacopa monnieri complexed with selected phospholipids and related substances can be used to significantly enhance cognitive performance beyond the levels provided by the non-complexed extract or by the extract formulated with conventional phospholipids. These effects have been tested and verified with respect to several diverse parameters relevant to cognitive performance. Specifically, the invention relates to a pharmaceutical or food composition comprising a Bacopa monnieri extract complexed with phosphatidylserine in the presence of one or more phosphoryl ether compounds such as phosphorylserine and phosphorylcholine. The compositions may further include complementary active ingredients and excipients for pharmaceutical and / or food use.The complexes according to the present invention can be used to enhance cognitive function and reduce mental fatigue in elderly people and children. Another aspect of the invention is the possibility of preventing cognitive decline. Yet another aspect of the present invention is the possibility of providing a drug or a food supplement for the treatment and prevention of pathological conditions related to reduced cognitive function and mental fatigue, such as dementia and Alzheimer's disease. Description of the drawings Figure 1: Effects of Bacopa extract in the following test: Numbers monitoring RT, Numbers monitoring accuracy, Choice RT accuracy. P = placebo; B = uncomplexed Bacopa extract; BP = Bacopa extract complexed with phosphatidylserine; BPP = Bacopa extract complexed with phosphatidylserine and phosphorylserine. Figure 2: Effects of Bacopa extract on the memory speed test: Number vigilance RT, Number vigilance accuracy, Choice RT accuracy. P = placebo; B = uncomplexed Bacopa extract; BP = Bacopa extract complexed with phosphatidylserine; BPP = Bacopa extract complexed with phosphatidylserine and phosphorylserine. Figure 3: Effects of Bacopa extract on the following tests: Numerical working memory RT; Word recognition RT; Spatial memory RT; Image recognition RT. P = placebo; B = uncomplexed Bacopa extract; BP = Bacopa extract complexed with phosphatidylserine; BPP = Bacopa extract complexed with phosphatidylserine and phosphorylserine. Figure 4: Effects of Bacopa extract on the memory quality test. P = placebo; B = uncomplexed Bacopa extract; BP = Bacopa extract complexed with phosphatidylserine; BPP = Bacopa extract complexed with phosphatidylserine and phosphorylserine. Figure 5: Effects of Bacopa extract on the image recognition accuracy test. P = placebo; B = uncomplexed Bacopa extract; BP = Bacopa extract complexed with phosphatidylserine; BPP = Bacopa extract complexed with phosphatidylserine and phosphorylserine. Figure 6: Effects of Bacopa extract on the mood test (calm). P = placebo; B = uncomplexed Bacopa extract; BP = Bacopa extract complexed with phosphatidylserine; BPP = Bacopa extract complexed with phosphatidylserine and phosphorylserine. Detailed description of the invention The present invention relates to novel complexes of Bacopa monnieri with phosphatidylserine (referred to herein as PTS) formed in the presence of at least one phosphoryl ether, such as phosphorylserine (referred to herein as PRS) and phosphorylcholine (referred to herein as PRC). It has been observed that simple complexes of Bacopa with phospholipids, for example, phosphatidylserine, are insufficient to achieve significant / systematic memory enhancement: they generally exhibit weak activity, sometimes indistinguishable from the group administered Bacopa alone or even placebo. In contrast, the selected formulation obtained from PTS in the presence of phosphoryl ether compounds, such as PRS and / or PRC, consistently showed very high performance, both in absolute and comparative terms, across a variety of qualitative and quantitative tests relevant to cognitive performance. Phosphatidylserine is a naturally occurring phospholipid defined by the structural formula (I) (YO) in which the R1 (C=O) O- and R2 (C=O) O- groups represent naturally occurring C3-30 fatty acid residues. The phosphoryl ether compound(s) used in the invention consist of a phosphoric acid group attached to an alkane, thus forming an ether linkage; the alkane may be further substituted with one or more amino and / or carboxyl groups. The preferred phosphoryl ethers for use in the invention are phosphorylserine and / or phosphorylcholine. The present formulation generally uses Bacopa monnieri in extract form. The term "extract" means that the plant or plant components are extracted with a suitable solvent well known in the prior art of plant extraction. According to the invention, the extract forms a complex with PTS in the presence of phosphoryl ether. The complex is formed by contacting the extract with a phospholipid mixture comprising: (a) PTS and (b) at least one phosphoryl ether. In a general embodiment, component (a) is present in an amount of at least 10%, more preferably at least 20%, and most preferably approximately 50% by weight of the total phospholipids in the mixture. In a secondary embodiment, both components (a) and (b) are present in an amount of at least 10%, more preferably at least 20%, and most preferably approximately 50% by weight of the total phospholipids in the mixture.The other phospholipids that make up the phospholipid mixture, besides PTS, if present, may be conveniently selected from commonly produced phospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, derivatives thereof, etc. Phosphoryl ethers are not phospholipids and are not counted in the "total phospholipids" referred to herein. In one embodiment, PTS is substantially the only phospholipid component of the phospholipid mixture, i.e., it constitutes up to 99.9% of the phospholipid mixture. In the manufacturing process of the present complexes, it is important that the extract comes into contact with PTS in the presence of phosphoryl ether (as opposed to independent contact of the extract with PTS followed by the addition of phosphoryl ether or vice versa): this ensures that the actual complexation of Bacopa with PTS takes place in the presence of phosphoryl ether, which has been found to be desirable for the efficacy of the formulation. The phospholipid mixture comprising PTS and phosphoryl ether is preferably dissolved in a suitable solvent and then contacted with a Bacopa monnieri extract at a mixture-to-extract ratio generally ranging from 1:1 to 6:1, preferably from 2:1 to 4:1, most preferably about 3:1. The suitable solvent can be selected from cyclic ethers, aromatic hydrocarbons, halogenated aromatic hydrocarbons, mixtures of alcohol with halogenated hydrocarbons (e.g., methanol:methylene chloride mixture), or aliphatic esters (e.g., ethyl acetate). The ratio of Bacopa extract to the phosphatidylserine used to prepare the complex is preferably from 1:3 to 1:6, most preferably about 1:5.The Bacopa extract is mixed with the phospholipid mixture, preferably with stirring, at a temperature between 30 and 100°C, or between 50 and 90°C, preferably around 70°C, until the solution is completely homogeneous. The complex can be recovered in solid form by evaporating the solvent and drying the residue, preferably at a temperature between 30 and 50°C for 24 to 48 hours. The formation of the complex can be determined by NMR; the resulting product exhibits physicochemical and spectroscopic characteristics that are markedly different from those of the original components and, as such, can be incorporated as an active ingredient in pharmaceutical formulations, particularly in oil-based formulations, which are preferable. The resulting complex is generally lipophilic and soluble in nonpolar and aprotic solvents. The complex shows significantly higher activity compared to free Bacopa monnieri extract, as well as to the extract complexed with PTS but without phosphoryl ether: it is suitable for incorporation into formulations where the extract alone cannot be used and leads to useful combinations. These complexes may be formulated as a pharmaceutical composition or food supplement, in combination with pharmaceutically or food-acceptable excipients and / or carriers. The formulation may be prepared using conventional methods, such as those described in the "Remingtons Pharmaceutical Handbook". The preferred carriers are oils rich in Q-3 fatty acids, as they facilitate the absorption of lipophilic complexes. Specifically, the carrier may comprise a combination of saturated and / or unsaturated fatty acids; the combination may consist of 15 to 25% by weight of saturated fatty acids, 5 to 15% by weight of monounsaturated fatty acids, and 60 to 80% by weight of polyunsaturated fatty acids, based on the total fatty acids in the combination. The composition is preferably formulated for oral administration and may be provided in any form suitable for this purpose, such as tablets, granules, powders, capsules, syrups, solutions, suspensions, coated tablets, gels, or eye drops. When formulated for other routes of administration, it may be provided, for example, as a solution or suspension for injection. The pharmaceutical composition or food supplement is provided in a form suitable for administering an effective daily amount of the present complex; the amount is preferably from approximately 50 to approximately 500 mg per day, particularly from approximately 100 to approximately 420 mg. According to a further aspect, the composition of the invention can be administered in combination with other substances that have a useful or complementary activity, such as extracts of Ginkgo biloba, Centella asiatica, Huperzia serrata, Olea europaea, Salvia officinalis, and drugs such as vinpocetine, galantamine, and donepezil. This composition may further include additional active ingredients and / or excipients; among the additional active ingredients are vitamins such as B1, B6, and B12, and / or amino acids such as acetylglutamine, asparagine, carnitine, and acetylcarnitine. Possible additional active ingredients include acetylcholinesterase inhibitors and complexed grape seed extracts. The complex according to the present invention significantly enhances cognitive performance, achieving numerous effects including: improved memory speed and quality, improved reaction time and working memory, prevention of memory decline, reduced mental fatigue, and a positive influence on mood and physical well-being, particularly in individuals with reduced cognitive function, such as middle-aged and elderly people. Therefore, a wide range of diverse abilities are positively influenced by these complexes, providing a comprehensive therapeutic approach to cognitive decline.Among the enhanced abilities, the following tests can be mentioned: Number vigilance time, number vigilance accuracy, choice time accuracy, memory speed, numerical working memory, word recognition time, spatial memory time, image recognition time, memory quality, image recognition accuracy, mood. The general context will become clear from the studies explained in the experimental part. Examples The activity of the combination of the invention is verified by methods described in the literature and based on a series of computerized evaluations (CDR). The series of computerized cognitive drug research assessments has been used in hundreds of clinical drug trials and has been shown to be sensitive to acute cognitive improvements as well as impairments with a wide variety of substances and botanical extracts. In this study, the additional task of rapid visual information processing (RVIP) was included in the series. The computer-controlled task selection from the system was administered by presenting parallel test forms in random order at each testing session. The presentation was conducted using desktop computers, and all responses were recorded using two-button (YES / NO) response boxes, apart from word presentation and immediate word recall. The other tests were: Image presentation (a series of 20 photographic images of everyday objects were presented on the monitor at a rate of 1 every 3 seconds, with a stimulus duration of 1 second, for the participant to remember them). Simple reaction time: Participants were instructed to press the "YES" response button as quickly as possible each time the word "YES" appeared on the monitor. Fifty stimuli were presented with an interval between stimuli that varied randomly between 1 and 3.5 seconds. Reaction times were recorded in milliseconds. Number monitoring task: A target number was randomly selected and continuously displayed on the right side of the monitor screen. A series of numbers was presented in the center of the screen at a rate of 80 per minute, and the participant was instructed to press the "YES" button as quickly as possible each time a number in the series matched the target number. The task lasted 15 minutes, and there were 15 stimulus-target matches. The task measures were accuracy (%), reaction time (ms), and number of false alarms. Choice reaction time: either the word "NO" or the word "YES" was presented on the monitor, and the participant was asked to press the corresponding button as quickly as possible. There were 50 trials, of which the 20 stimulus words were chosen randomly with equal probability, with a randomly varying interval between stimuli of between 1 and 3.5 seconds. Reaction times (ms) and accuracy (%) were recorded. Spatial working memory: A graphical representation of a house was presented on the screen with four of its nine windows illuminated. The participant was instructed to memorize the position of the illuminated windows. In 36 subsequent presentations of the house, one of the windows was illuminated, and the participant decided whether or not it matched one of the illuminated windows in the original presentation. The participant gave their response by pressing the "YES" or "NO" response button as quickly as possible. Mean reaction times were measured in ms, and the accuracy of responses to both original and novel (distracting) stimuli was recorded as percentages, which were used to derive a "% greater than chance performance" score (percentage of 5 original targets + percentage of novel targets correctly identified - 100). Numerical working memory: Five numbers were presented sequentially for the participant to recall. This was followed by a series of 30 probe numbers, for each of which the participant decided whether it had been in the original series and pressed the "YES" or "NO" response button as appropriate as quickly as possible. This was repeated two more times with different stimuli and probe numbers. Mean reaction times were measured in milliseconds, and the accuracy of responses to both original and novel (distracting) stimuli was recorded as percentages, which were used to derive a "% greater than chance performance" score, as described above. Delayed word recall: Participants were again given 60 seconds to write down as many of the words as possible. The task was scored for the number of correct words, and the resulting score was converted into a percentage. Delayed word recognition: The original words plus 15 distraction words were presented one at a time in a randomized order. For each word, the participant indicated whether they recognized it as being included in the original word list by pressing the "YES" or "NO" button as appropriate and as quickly as possible. Mean reaction times were measured in ms, and the accuracy of responses to both original and novel (distraction) stimuli was recorded as percentages, which were used to derive a "% greater than 25 chance performance" score as before. Delayed image recognition: The original images plus 20 distractor images were presented one at a time in a randomized order. For each image, participants indicated whether they recognized it as belonging to the original series by pressing the "YES" or "NO" button as appropriate and as quickly as possible. Mean reaction times were measured in milliseconds, and the accuracy of responses to both original and novel (distractor) stimuli was recorded as percentages, which were used to derive a "% greater than chance performance" score as before. Primary cognitive outcome measures The above measures were combined to yield a "memory quality" measure and five serially derived cognitive outcome factors. These measures were used in several studies, including a repeated assessment of the cognitive effects of herbal medicines. Attention "Attention speed" factor: derived by combining the reaction times of the three attention tasks: simple reaction time, choice reaction time, and number vigilance (units are milliseconds summed for the 3 tasks). "Attention accuracy" factor: derived by calculating the combined percentage accuracy across the 20 number choice and number vigilance reaction time tasks, adjusted for false alarms in the latter test. 100% accuracy across both tasks will result in a maximum score of 100. Memory "Memory speed" factor: derived by combining the reaction times of the four computerized memory tasks: numerical working memory, spatial memory, delayed word recognition, and delayed image recognition (units are milliseconds summed for the 4 tasks). Measure of "memory quality": derived by combining the "secondary memory" and "working memory" factor scores (see below). "Secondary memory" factor: derived by combining the percentage accuracy scores (adjusted for proportions of novel and original stimuli where appropriate) from all other secondary memory tests: delayed word recognition, delayed picture recognition, immediate word recall, and delayed word recall (with adjustments for the total percentage correct for errors and intrusions in the latter two tasks). One hundred percent accuracy across all four tasks will generate a maximum score of 400 on this index. Working memory factor: derived by combining the percentage accuracy scores from the two working memory tests: spatial working memory and numerical working memory. One hundred percent accuracy across both tasks will generate a maximum score of 200 on this index. Subtraction tasks involving 3 and 7 in series A modified computerized version of the serial sevens test was used. The original verbal serial sevens test has been described in various ways, including as part of the Mini-Mental State Examination. It has been used, for example, to assess cognitive impairment during hypoglycemia and to investigate the relationship between elevated blood glucose levels and cognitive performance. In the current studies, computerized versions of serial subtraction were implemented, using 2-minute tests. For the serial sevens section, a conventional instruction screen instructed the participant to count backward by sevens from the given number, as quickly and accurately as possible, using the numeric keypad to enter each response. Participants were also verbally instructed that if they made a mistake, they should continue subtracting from the new incorrect number.A random initial number between 800 and 999 was displayed on the computer screen and cleared after the first response. Each three-digit answer was entered using the numeric keypad, with each digit represented on the screen by an asterisk. Pressing the Enter key signaled the end of each response and cleared the three asterisks from the screen. The task was scored based on the total number of subtractions and the number of errors. In the case of incorrect answers, subsequent correct answers were scored positively. The serial threes task was identical to the serial sevens task, except that it involved serial subtraction of threes. Measure of subjective mood Bond-Lader visual analogue scales, consisting of 16 100mm visual analogue scales anchored by antonyms (e.g., alert-sleepy, lethargic-energetic, etc.) as recommended by the authors, were combined to form three mood factors: alertness, tranquility, and contentment. Treatments Participants Twenty-eight female and fifteen male students (mean age 22.6 years) participated in the study after approval by the ethics committee and after signing an informed consent form and completing a medical health questionnaire. All participants reported being in good health, not using any recreational or over-the-counter drugs, and very familiar with using personal computers. Regular smokers or students who consumed alcohol were excluded. On each day of the study, participants received two capsules that looked identical each time. The individual capsules contained: - an inert placebo, or -100 mg of Bacopa monnieri extract (hereinafter referred to as "B"), or - 300 mg of Bacopa monnieri extract complexed with a phospholipid mixture containing 40% phosphatidylserine (equivalent to 100 mg of Bacopa monnieri extract), hereinafter referred to as "BP", or - 300 mg of Bacopa monnieri extract complexed with a phospholipid mixture containing 20% ​​phosphatidylserine, supplemented with 20% phosphorylserine (equivalent to 100 mg of Bacopa monnieri extract) and hereafter referred to as "BPP". To maintain double-blinding, manufacturer-coded treatments were provided in identical hard gelatin capsules. A third, unbiased party was then responsible for preparing treatments according to the study's Latin square. The code remained unbroken until the initial statistical analysis was completed. All treatments looked and smelled the same. Procedure Each participant was required to attend a total of five days of the study, which were conducted seven days apart to ensure sufficient washout time between conditions. The tests took place, starting at the same time each day, in a series of laboratories with participants visually isolated from one another. Upon arrival at their first session on day one, participants were randomly assigned to a treatment regimen using a Latin square design that compensated for the order of treatments across the four active days of the study. Day one was identical to the following four, except that no treatment (active or placebo) was offered, to allow for familiarization with the test series and procedure. Data from the four practice sessions on this day were not included in any analyses. Each day of the active study comprised four identical testing sessions. The first was a pre-dose testing session that established baseline performance for that day and was immediately followed by the 20-day treatment on days 2 through 5. Additional testing sessions began at 1, 3, and 6 hours post-treatment consumption.Each testing session involved completing the Bond-Lader visual analog scales and the CDR test series. Statistical data Scores on individual task outcomes, the four primary factors, and the two secondary memory factors were analyzed as "change from baseline" using the Minitab statistical package. Before conducting planned comparisons, an ANOVA (general linear model), with model-fitted terms for dose, visit, dose x visit, and subject, was performed to identify main and interaction effects on change-from-baseline data for each measure. Primary statistical analysis of the "change-from-baseline" data for each measure was performed using planned comparisons, employing mean square t-tests for "dose * time * subjects" derived from an omnibus ANOVA as the error term. At each time point (1, 2, 5, 4, and 6 hours post-dose), data from the placebo condition were compared with those from each of the three treatments (B, BP, BPP).To ensure the level of protection against overall Type I error, only planned comparisons associated with measures that generated a significant main effect or interaction effect in the initial ANOVA are reported. Furthermore, all tests were two-sided, comparisons were strictly planned before the study, the number of conditions was limited to at least 15 (one at each time point), and only probabilities associated with these pre-planned comparisons were calculated. Results Overall, the Bacopa monnieri extract complex with phosphatidylserine plus phosphorylserine according to the present invention (BPP) had generally superior efficacy compared to the other species tested, i.e., placebo (P), non-complexed Bacopa extract (B), or Bacopa complex with phosphatidylserine (BP), with respect to all parameters tested. The results are reported graphically in Figures 1 to 6 and are summarized comparatively in the following Table 1 in which, for each of the tests performed, the number of asterisks defines the average effect size in the group of participants treated according to the invention (BPP) during the entire testing period, compared to the reference groups (P, B and BP). Table 1 It appears that, in each test, the BPP group scored better than the corresponding reference groups. In particular, the difference with the other group tested was most notable in the following tests: memory speed, numerical working memory, word recognition, and image recognition accuracy.

Claims

1. A complex of Bacopa monnieri extract with a phospholipid mixture comprising phosphatidylserine, the complex being formed in the presence of a phosphoryl ether.

2. The complex according to claim 1, wherein the phosphoryl ether is selected from phosphorylserine and / or phosphorylcholine.

3. The complex according to claims 1-2, wherein said phosphatidylserine is used in an amount of at least 10%, preferably at least 20%, more preferably approximately 50% by weight relative to the total phospholipids of said phospholipid mixture.

4. The complex according to claims 1-3, wherein said phosphoryl ether is used in an amount of at least 10%, preferably at least 20%, more preferably approximately 50% by weight relative to the total phospholipids of said phospholipid mixture. 5.A complex according to claims 1-4, wherein the phospholipid mixture further contains one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, and derivatives thereof.

6. A complex according to claims 1-5, wherein the weight ratio of Bacopa extract to the phosphatidylserine used to prepare the complex is from 1:3 to 1:

6.

7. A process for preparing a complex of Bacopa monnieri extract with a phospholipid mixture according to claim 1, comprising: a) providing a phospholipid mixture comprising phosphatidylserine, supplemented with a phosphoryl ether; b) mixing the product of a) with a Bacopa monnieri extract.

8. A process according to claim 7, wherein said phosphoryl ether is selected from phosphorylserine and / or phosphorylcholine. 9.A process according to claims 7-8, wherein said phosphatidylserine is used in an amount of at least 10%, preferably at least 20%, more preferably approximately 50% by weight relative to the total phospholipids of said phospholipid mixture.

10. A process according to claims 7-9, wherein said phosphoryl ether is used in an amount of at least 10%, preferably at least 20%, more preferably approximately 50% by weight relative to the total phospholipids of said phospholipid mixture.

11. A process according to claims 7-10, wherein the phospholipid mixture further contains one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, and derivatives thereof. 12.A process according to claims 7-11, wherein the product of step a) is mixed with Bacopa monnieri extract in a weight ratio ranging from 1:1 to 6:1, preferably from 2:1 to 4:

1.

13. A process according to claims 7-12, wherein the product of step a) is mixed with Bacopa monnieri extract with stirring, at a temperature from 30 to 100°C, preferably from 50 to 90°C.

14. A pharmaceutical composition or food supplement comprising the complex according to claims 1-6 in combination with pharmaceutical or food carriers / excipients.

15. A pharmaceutical composition or food supplement according to claim 14, further comprising other active ingredients that are effective in enhancing memory. 16.A pharmaceutical composition or food supplement according to claims 14 and 15, wherein said other active ingredients are selected from: Ginkgo biloba, Centella asiatica, Huperzia serrata, Olea europaea, Salvia officinalis, vinpocetine, galantamine, donepezil; vitamins such as B1, B6, and B12; amino acids such as acetylglutamine, asparagine, carnitine, and acetylcarnitine; acetylcholinesterase inhibitors; and complexed grape seed extracts.

17. A pharmaceutical composition or food supplement according to claims 14 and 16 for use in a method of enhancing memory and cognitive functions.

18. A pharmaceutical composition or food supplement for use according to claim 17, for reducing mental fatigue, improving memory speed, improving memory quality, preventing or reducing cognitive impairment, reversing drug- or stress-related amnesic effects, and treating memory deficits in Alzheimer's disease or other forms of dementia.