COMPOSITIONS AND METHODS FOR NASAL ADMINISTRATION OF DRUGS TO BRAIN AND FOR SYSTEMIC EFFECT

Abstract

The invention relates to a nasally administrable composition comprising at least one active substance in magnesium-containing vesicular carrier, said carrier comprising glycol, phospholipids, water and at least one magnesium source. Methods for nasal administration of the composition, for example, for pain relief, are also provided.

Claims

1-19. (canceled)

20. A method of administering an active substance to a mammal in need thereof, comprising intranasal administration of a composition comprising a therapeutically effective amount of the active substance in a magnesium-containing vesicular carrier that contains glycol, phospholipids, water and at least one magnesium source.

21. A method according to claim 20, wherein the composition further comprises an antioxidant, a mucoadhesive agent or both.

22. A method according to claim 20, wherein the composition is free of aliphatic monohydroxy alcohol.

23. A method according to claim 20, wherein the active substance is selected from the group consisting of an analgesic, antimigraine and/or antipyretic agents; opioids; an anti-Parkinson drug; a sedative and/or hypnotic drug.

24. A method according to claim 20, wherein the active substance is selected from the group consisting of peptide or polypeptides.

25. A method of increasing the delivery of a physiologically active compound from a nasally administrable composition to the bloodstream/brain of mammals, the method comprises incorporating magnesium source into a vesicular carrier which contains glycol, phospholipids, water and said physiologically active compound, for enhanced delivery.

26. A method according to claim 25, wherein the active compound is selected from the group consisting of an analgesic, antimigraine and/or antipyretic agents; an opioid; an anti-Parkinson drug; a sedative and/or hypnotic drug; and a hormone peptide.

27. (canceled)

28. A method of treating Parkinson's disease, symptoms associated with Parkinson's disease or Parkinsonism, which method comprises administering intranasally, to a patient in need thereof, a pharmaceutical composition comprising pramipexole and a vesicular carrier.

29. A method according to claim 28, for treating impaired locomotion in Parkinson's disease patients.

30. A method according to claim 28, wherein the carrier is a magnesium-containing vesicular carrier comprising glycol, phospholipids, water and at least one magnesium source.

31-33. (canceled)

Description

[0134] In the drawings:

[0135] FIG. 1: Three-dimensional micrographs of the olfactory region for mice brain treated with R6G at a dose of 10 mg/kg from phospholipids magnesome, water solution and Liposome. Height: 589 μm, width: 589 μm, depth: 606 μm, lens ×20 (Al-MP microscope NIKON—Japan).

[0136] FIG. 2: a bar diagram showing fluorescent intensity (A.U.) in the olfactory region of mice brain at 10 min after treatment with R6G at a dose of 3 mg/kg in phospholipids magnesome, water solution and Liposome; Mean±SD. Auto fluorescence was subtracted. **P value <0.01 considered very significant.

[0137] FIG. 3: Three-dimensional micrographs of the olfactory region for mice brain treated with R6G at a dose of 10 mg/kg from phospholipid magnesome vs. soft vesicles. Height: 979 μm, width: 979 μm, depth: 507 μm, lens ×20 (Al-MP microscope NIKON-Japan).

[0138] FIG. 4: a bar diagram showing fluorescent intensity (A.U.) in the olfactory region of mice brain at 10 min after treatment with Insulin-FITC at a dose of 1 mg/kg in phospholipids magnesome, WS and Lipo; Mean±SD. Auto fluorescence was subtracted. **P value <0.01 considered very significant.

[0139] FIG. 5: NIR images for mice brains treated with EPG IRDye 800CW at a dose 1 mg/Kg from phospholipid magnesome as compared to water solution and liposome.

[0140] FIG. 6: a bar diagram showing fluorescent intensity (A.U.) in the olfactory region of mice brain at 10 min after treatment with EGF-IRDye 800CW at a dose of 1 mg/kg in phospholipids magnesome, as compared to water solution and liposome; Mean±SD. Auto fluorescence was subtracted. **P value <0.01 considered very significant.

[0141] FIG. 7: a bar diagram showing the writhing counts for mice treated with Oxytocin at a dose of 0.4 mg/kg phospholipid magnesome, WS, Lipo and Untreated control 5, 30 and 120 min prior to IP injection of acetic acid; Mean±SD. P<0.05 for phospholipid magnesome vs. untreated, WS and Lipo 5 min. P<0.01 for phospholipid magnesome vs. untreated, WS and Lipo at 30 and 120 min time points.

[0142] FIGS. 8A-8D: Representative micrographs of nasal cavities excised from rats that (A) received no treatment or treated with (B) phospholipid Magnesome, (C) NS and (D) SLS nasal solution.

[0143] FIGS. 9A-9B: DSC thermograms for (A) phospholipids magnesome, (B) liposome obtained by Mettler Toledo DSC-1 STAR system (China).

[0144] FIG. 10: a graph showing the pharmacokinetic profile of Ketoprofen in plasma following nasal administration of Ketoprofen from the composition of the invention vs. Ketoprofen oral administration, each at a dose of 14 mg/Kg. Results (mean±SD)** P<0.01, very significant at 10 min, and 30 min.

[0145] FIG. 11: Number of squares crossed in the open field test by Parkinson's mice treated with: Nasal Pramipexole Phospholipid Magnesome (n=6); Oral Pramipexole solution (n=4) and untreated animals (n=5), (Mean±SD). p<0.001 for Pramipexole Phospholipid Magnesome vs. untreated control, p<0.01 for Pramipexole Phospholipid Magnesome vs. oral, p>0.05 (considered not significant) for Pramipexole oral vs. untreated control by one-way ANOVA.

[0146] FIG. 12: Ptosis scores for Parkinson's mice model treated with: Nasal Pramipexole Phospholipid Magnesome (n=6); Oral Pramipexole Solution (n=4) and untreated animals (n=5), (Mean±SD). p<0.01 for Pramipexole Phospholipid Magnesome vs. untreated control, p>0.05 for Pramipexole nasal vs. oral and oral vs. untreated, by one-way ANOVA.

[0147] FIG. 13: Number of squares crossed in the open field test by Parkinson's mice model treated with: Nasal Pramipexole Phospholipid Magnesome; Oral Pramipexole solution and untreated animals (n=7/group), (Mean±SD). p<0.001 for Pramipexole Phospholipid Magnesome vs. untreated control, p<0.01 for Pramipexole Phospholipid Magnesome vs. oral, p>0.05 (considered not significant) for Pramipexole oral vs. untreated control by one-way ANOVA.

[0148] FIG. 14. Catalepsy duration measured by bar test for Parkinson's mice model treated with: Nasal Pramipexole Phospholipid Magnesome; Oral Pramipexole solution and □ untreated animals (n=7/group), (Mean±SD). p<0.001 for Pramipexole Phospholipid Magnesome vs. untreated control, p<0.01 for Pramipexole Phospholipid Magnesome vs. oral, p>0.05 (considered not significant) for Pramipexole oral vs. untreated control by two tail Mann-Whitney test.

[0149] FIG. 15. Ptosis scores for Parkinson's mice model treated with: Nasal Pramipexole Phospholipid Magnesome; Oral Pramipexole solution and untreated animals (n=7/group), (Mean±SD). p<0.001 for Pramipexole Phospholipid Magnesome vs. untreated control, p<0.01 for nasal vs. oral and p>0.05 for oral vs. untreated, by one-way ANOVA.

EXAMPLES

Abbreviations Used in the Examples

[0150] PL—Phospholipid

[0151] PG—Propylene Glycol

[0152] WS—Water solution

[0153] DDW—Double distilled water

[0154] ETOH—Ethanol

[0155] HSO—Hemp Seed Oil

[0156] SO—Sesame oil

[0157] Lipo—Liposome

[0158] The following materials were used: Magnesium Sulfate, anhydrous from J.T.Baker, USA; Phospholipon 90 G—Lipoid, Phospholipid GmbH, Germany; Propylene Glycol—Tamar, Israel and Vitamin E Acetate—Tamar, Israel.

Example 1

Nasal Delivery to Brain

[0159] Nasal delivery of R6G in the composition of the invention to the olfactory region of mice brain visualized by a Multiphoton Microscope (Al-MP microscope NIKON—Japan).

[0160] Part I

[0161] Nine female C57Bl/6J mice (8-9 weeks), were divided equally into 3 groups of administration. The following formulations were prepared:

TABLE-US-00001 Phospholipid Water magnesome solution Liposome (% w/w) % w/w % w/w R6G 1 1 1 PL 3 — 3 PG 15 — — Sodium alginate 0.6 — 0.6 Magnesium sulfate 0.03 — — DDW To 100 To 100 To 100

[0162] Preparation: PL was dissolved in PG, R6G was added and then 20% of the total DDW amount was added, through mixing. In a separate vessel, Sodium Alginate was dispersed in 10% of the total amount of DDW and mixed with Magnesium Sulfate aqueous solution (10 mg/ml). The PL solution was then added to the alginate gel through mixing with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany), then the remaining DDW was added.

[0163] R6G was administered nasally to mice from the three above formulations at a dose of 10 mg/kg animal (˜0.2 mg\ 20 μl/mouse). Ten minutes after treatments, the animals were sacrificed, the brains were removed, washed with normal saline and examined by the Multiphoton Microscope using the following conditions: excitation λ 850 nm, field of image of 589×589×606 nm (width×height×depth), lens ×60 laser, intensity 5%, scanning 512, scan speed 0.5, line skipping 2, Luts1500 and zoom1. The fluorescence intensity of the probe (arbitrary units A.U.) in the olfactory region in brain was further assessed using Image Pro-Plus software.

[0164] The micrographs are presented in FIG. 1 (left: phospholipid magnesome; middle: water solution; right: liposome). FIG. 2 is a bar diagram showing the fluorescent intensity. The micrographs of Multiphoton Microscopic examination and the semi-quantification showed that higher fluorescent signals are found in the olfactory region in the groups treated with phospholipid magnesome containing R6G relative to different control nasal compositions Water Solution (WS) and Liposome (Lipo). It is worthy to notice that the fluorescence intensity in the WS group was apparently higher (yet not statistically significant) than that achieved in the animals receiving Lipo treatment.

[0165] These results point towards the efficiency of phospholipid magnesome to improve the hydrophilic probe delivery to the examined region in the brain.

[0166] Part II

[0167] In this part, the effect of phospholipid magnesome was evaluated in comparison with a composition without magnesium.

[0168] Female C57Bl/6J mice (8-9 weeks), were divided into 2 groups of administration. The following formulations were prepared:

TABLE-US-00002 Phospholipid No Mg magnesome (% w/w) % w/w R6G 1 1 PL 3 3 PG 15 15 Magnesium sulfate 0.03 — DDW To 100 To 100

[0169] The administered dose and the experimental procedure was performed as described in Part I of this example.

[0170] FIG. 3 shows the three-dimensional micrographs. The results of this part of the experiment show deeper delivery of R6G into the examined brain region. The semi quantification indicated a fluorescence of 13.2 A.U. to the phospholipid magnesome as compared to 8.0 A.U. for the soft vesicles, such results pointing towards the superiority of phospholipid magnesome as carrier for brain delivery of drugs.

Example 2

Nasal Delivery of Insulin-FITC to Brain

[0171] Nasal delivery of Insulin FITC to the olfactory region in mice brain from the composition of the invention and two control compositions, examined by a Multiphoton Microscope (Al-MP microscope NIKON—Japan).

[0172] Nine female C57Bl/6J mice (8-9 weeks) were divided equally into groups of administration. The following formulations were prepared.

TABLE-US-00003 Phospholipid Water magnesome solution Liposome (% w/w) % w/w % w/w Insulin FITC 0.1 0.1 0.1 PL 3 — 3 PG 15 — — Sodium alginate 0.6 — 0.6 Magnesium sulfate 0.03 — — DDW To 100 To 100 To 100

[0173] Preparation: PL was dissolved in PG, Insulin FITC was added then 20% of the total DDW amount was added, through mixing. In a separate vessel, Sodium Alginate was dispersed in 10% of the total amount of DDW and mixed with Magnesium Sulfate aqueous solution (10 mg/ml). The PL solution was then added to the alginate gel through mixing and then the remaining DDW was added.

[0174] Insulin FITC was administered nasally to mice from the three formulations at a dose of 1 mg/kg animal (˜0.02 mg\ 20 μl/mouse). Ten minutes after treatments, the animals were sacrificed, the brains were removed, washed with normal saline and examined by the Multiphoton Microscope using the following conditions: excitation λ of 860 nm, field 58.6×58.6×30.6 nm (width×height×depth), lens ×20, laser intensity 11.1%, scanning 512, scan speed 0.5, no line skipping, Luts: 1000 and zoom 9.9. The fluorescence intensity of the probe (arbitrary units A.U.) in the olfactory region in brain was further assessed using Image Pro-Plus software.

[0175] Multiphoton imaging of the olfactory region following Insulin-FITC administration in phospholipid magnesome indicates the presence of augmented fluorescent signal in this group relative to controls (micrographs are not shown). Semi-quantification of the fluorescence signals gave fluorescent intensities of ˜24 A.U. in brain section of animals treated with phospholipid magnesome containing Insulin-FITC as compared to 3.4 and 7.0 A.U. for the controls WS and Lipo, respectively, as shown in the bar diagram of FIG. 4.

Example 3

Nasal Delivery of EGF-IRDye 800CW to Brain

[0176] Nasal delivery of EGF-IRDye 800CW to brain of mice from phospholipid magnesome as compared with two controls, was examined by Odyssey® Infrared Imaging System (LI-COR, USA).

[0177] Twelve female C57Bl/6J mice (8-9 weeks) were divided equally into 3 groups of administration and Untreated Control group. The following formulations were prepared.

TABLE-US-00004 Phospholipid Water magnesome solution Liposome (% w/w) % w/w % w/w EGF-IRDye 800CW 0.1 0.1 0.1 PL 3 — 3 PG 15 — — Sodium alginate 0.6 — 0.6 Magnesium sulfate 0.03 — — DDW To 100 To 100 To 100

[0178] Preparation: PL was dissolved in PG, EGF-IRDye 800CW was added and then 20% of the total DDW amount was added, through mixing. In a separate vessel, Sodium Alginate was dispersed in 10% of the total amount of DDW and mixed with Magnesium Sulfate aqueous solution (10 mg/ml). The PL solution was then added to the alginate gel through mixing, then the remaining DDW was added.

[0179] EGF-IRDye 800CW was administered nasally to mice from the three formulations at a dose of 1 mg/kg animal (˜0.02 mg\ 20 μl/mouse). Ten minutes after treatments, the animals were sacrificed; brains were removed, washed with normal saline and observed under the imaging system. The scanning was performed using offset 3, resolution 339.6 μm, channel 800 nm and intensity 3.

[0180] The NIR images (FIG. 5; from left to right: untreated group, phospholipid magnesome-treated group; water solution-treated group and liposome-treated group) and their semi-quantification (FIG. 6, in the form of a bar diagram) indicate the presence of the labeled peptide signal in brain tissues following nasal administration from various systems as compared to Untreated Control. FIG. 5 shows that following administration from phospholipid magnesome, EGF-IRDye 800CW accumulated in the cerebrum and in the olfactory bulb with remarkable accumulation in the cerebrum. The administration from water solution lead to peptide accumulation in the olfactory bulb (FIG. 5). The fluorescent signals calculated to be 19.1 A.U. in phospholipid magnesome containing EGF-IRDye 800CW as compared to 10.0 and 6.7 A.U. in WS and Lipo, respectively (FIG. 6).

Example 4

Analgesic Effect of Oxytocin Nasally Delivered in Phospholipid Magnesome as Compared to Two Control Carriers

[0181] The analgesic effect of intranasal administration of Oxytocin in phospholipid magnesome composition was evaluated in female C57Bl/6J mice. The acetic acid-induced pain mice model was used.

[0182] In this model, animals received analgesic treatment, then after predetermined time periods, pain was induced by 0.6% (v/v) acetic acid solution injected intraperitoneally at a dose of 10 ml/kg. The number of writhes in a 10 min period was counted, starting 5 min after the acetic acid injection. A writhe is characterized by a wave of contraction of the abdominal musculature followed by extension of at least one hind limb. Antinociception is expressed as percent inhibition of the number of writhes observed in treated animals in comparison to animals in the untreated group.

[0183] The Maximum Possible Effect (MPE %) of different treatments is expressed as the inhibition percent of the number of writhes in a drug-treated animal group, when compared to the mean number of writhes measured in a group of untreated control mice according to the following equation:

MPE %=[Mean of writhes in untreated control group−Mean of writhes in treated group]/[Mean of writhes in untreated control group]*100

[0184] Twenty mice were divided into three equal treatment groups for testing three time points: 5, 30 or 120 min (n=5/group) and Untreated control group. The following formulations were prepared.

TABLE-US-00005 Phospholipid Water magnesome solution Liposome (% w/w) % w/w % w/w oxytocin 0.08 0.08 0.08 PL 3 — 3 PG 15 — — Sodium alginate 0.6 — 0.6 Magnesium sulfate 0.03 — — DDW To 100 To 100 To 100

[0185] Preparation: PL was dissolved in PG, Oxytocin was added and then 20% of the total DDW amount was added, through mixing. In a separate vessel, Sodium Alginate was dispersed in 10% of the total amount of DDW and mixed with Magnesium Sulfate aqueous solution (10 mg/ml). The PL solution was then added to the alginate gel through mixing, then the remaining DDW was added.

[0186] Oxytocin was administered nasally to mice from the three formulations at a dose of 0.4 mg/kg animal (˜0.008 mg\ 10 μl/mouse).

[0187] FIG. 7 and the table below give the writhes counts and MPE % values, respectively, following treatment of mice with 0.4 mg/kg Oxytocin incorporated in various carriers. As shown in the bar diagram of FIG. 7, a significant decrease in writhes number was observed following treatment with Oxytocin from phospholipid magnesome (the darkest bar). The effect of the peptide incorporated in various controls (WS and Lipo) was lower by 2 folds, underscoring the enhanced delivery of Oxytocin from phospholipid magnesome.

[0188] Below are tabulated calculated MPE % values following Oxytocin administration to mice at a dose of 0.4 mg/kg from phospholipid magnesome and control nasal systems administrated at various time points before pain induction.

TABLE-US-00006 MPE % Phospholipid Water Time (min) magnesome solution Liposome 5 63.5 35.8 30.8 30 63.9 27.8 25.8 120 58.0 29.8 19.2

Example 5

Local Safety

[0189] The effect of phospholipid magnesome on the nasal cavity was evaluated in rats. Female SD/H rats were divided into four groups:

[0190] Group 1—(phospholipid magnesome): Intranasal administration of phospholipid magensome composition as described in Example 1,

[0191] Part II (15 μl/rat)

[0192] Group 2—(NS): Intranasal Normal saline (15 μl/rat).

[0193] Group 3—(SLS): Intranasal Sodium lauryl sulfate Solution (1% w/w), (15 μl/rat).

[0194] Group 4—Untreated control animals.

[0195] The animals received the treatments twice a day for one week. At the end of the experiment, animals were sacrificed, nasal cavities were removed and fixed in 3.7% Formaldehyde PBS. Sections of the nasal cavity were cut serially at 7 μm thickness and stained with Hematoxylin & Eosin. The sections were examined by professional histopathologist (Authority for Animal Facilities, Hebrew University of Jerusalem, Israel) by Olympus light microscope BX43 and Olympus digital camera DP21 with Olympus cellSens Entry 1.13 software (Olympus, Japan) using magnification ×10. Local toxicity was assessed by evaluating the histopathological alterations in different regions of the nasal cavity (cartilage and turbinate bone, lamina propria and submucosa, mucosal epithelium and lumen).

[0196] No pathological findings were observed in the histopathological analysis of the nasal cavities excised from rats treated with phospholipid magnesome or NS. The micrographs for these groups were similar to untreated control group showing intact mucosal epithelium, empty lumen and no infiltration of inflammatory cells. Overall, there was no evidence of inflammation. Turbinate bone integrity was preserved. Epithelium was normal with no evidence of erosion or ulceration and ciliated epithelium was intact. On the other hand, minimal proteinaceous material in the lumen and focal aggregations of neutrophils were observed in the positive control group treated with SLS. Micrographs corresponding to the four groups are presented in FIG. 8.

Example 6

Tramadol HCl-Containing Phospholipid Magnesome

[0197]

TABLE-US-00007 Ingredients % w/w Tramadol HCl 10 PL 3 PG 15 Sodium Alginate 0.6 Magnesium Sulfate 0.01 DDW To 100

[0198] Preparation: PL was dissolved in PG, Tramadol and 20% of the total DDW amount were added, through mixing. In a separate vessel, Sodium Alginate was dispersed in 10% of the total amount of DDW and mixed with Magnesium Sulfate aqueous solution (10 mg/ml). The PL solution was then added to the alginate gel through mixing with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany), then the remaining DDW was added.

Example 7

Rizatriptan Benzoate-Containing Phospholipid Magnesome

[0199]

TABLE-US-00008 Ingredients % w/w Rizatriptan Benzoate 10 PL 3 PG 15 Magnesium Sulfate 0.03 DDW To 100

[0200] Preparation: PL was dissolved in PG, then Rizatriptan was added, through mixing. To this mixture, Magnesium Sulfate aqueous solution (10 mg/ml) was added through mixing with a magnetic stirrer, then the remaining DDW was added.

Example 8

Mannitol-Containing Phospholipid Magnesome

[0201]

TABLE-US-00009 Ingredients % w/w Mannitol 10 PL 3 PG 15 Magnesium Sulfate 0.07 DDW To 100

[0202] Preparation: PL was dissolved in PG, then Mannitol was dispersed in the PL solution. To this mixture Magnesium Sulfate aqueous solution (10 mg/ml) was added through mixing with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany), then DDW was added.

Example 9

Cannabidiol-Containing Phospholipid Magnesome

[0203]

TABLE-US-00010 Ingredients % w/w CBD 1 PL 3 PG 25 Ethanol Absolute (ETOH) 15 Magnesium Sulfate 0.01 DDW To 100

[0204] Preparation: PL was dissolved in ETCH and PG mixture, then CBD was dissolved in the PL solution. To this solution, Magnesium Sulfate aqueous solution (10 mg/ml) was added through mixing with a magnetic stirrer then DDW was added.

Example 10

Cannabidiol-Containing Phospholipid Magnesome

[0205]

TABLE-US-00011 Ingredients % w/w CBD 0.5 PL 5 PG 25 ETOH 17 Magnesium Sulfate 0.01 DDW To 100

[0206] The composition is prepared with a high shear mixer. PL was dissolved in ETOH and PG mixture, then CBD was dissolved in the PL solution. To this solution, Magnesium Sulfate aqueous solution (10 mg/ml) was added through mixing with a magnetic stirrer then DDW was added.

Example 11

Brotizolam-Containing Phospholipid Magnesome

[0207]

TABLE-US-00012 Ingredients % w/w Brotizolam 0.1 PL 3 PG 15 Carbopol 980 0.05 Ammonium hydroxide 0.05 Magnesium Sulfate 0.02 DDW To 100

[0208] PL was dissolved in PG. Brotizolam was added to the solution. In a separate vessel, Carbopol 980 was suspended in DDW and ammonium hydroxide was added. Then Magnesium Sulfate aqueous solution (10 mg/ml) was added to this mixture followed by adding the PL solution through mixing with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany).

Example 12

Cannabidiol-Containing Phospholipid Magnesome

[0209]

TABLE-US-00013 Ingredients % w/w CBD 1 PL 10 PG 40 Vit E 0.4 Magnesium Sulfate 0.03 DDW To 100

[0210] PL was dissolved in PG, then CBD and Vit E were dissolved in the PL solution. To this solution, Magnesium Sulfate aqueous solution (10 mg/ml) was added through mixing with a magnetic stirrer then DDW was added.

Example 13

Butorphanol Tartrate (BUT)-Containing Phospholipid Magnesome

[0211]

TABLE-US-00014 Ingredients % w/w BUT 0.1 PL 2 PG 30 Sodium Hydroxide 0.5 Magnesium Sulfate 0.025 DDW To 100

[0212] Preparation: PL was dissolved in PG, then BUT was dissolved in the PL solution. To this solution, DDW and magnesium sulfate solutions were added through mixing with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany). Finally, Sodium Hydroxide was added (for pH adjustment). Final pH—5.5.

Example 14

Ketoprofen (KET)-Containing Phospholipid Magnesome

[0213]

TABLE-US-00015 Ingredients % w/w KET 20 PL 2 Magnesium Sulfate 0.025 PG 30 Sodium Hydroxide (10%) 40 Hydrochloric Acid (32%) 5.8 DDW To 100

[0214] Preparation: PL was dissolved in PG. In a separate vessel, KET was suspended in 20% of DDW, then Sodium Hydroxide Solution was added (to dissolve KET) and followed by the addition of Hydrochloric Acid (for pH adjustment) and the rest amount of DDW and magnesium sulfate solution wee added. Finally, the PL solution was added to this solution and mixed with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany).

Example 15

Ketoprofen (KET)-Containing Phospholipid Magnesome

[0215]

TABLE-US-00016 Ingredients % w/w KET 3 PL 2 PG 30 Sodium Hydroxide (10%) 10 Magnesium sulfate 0.02 HCl solution (32%) 2 DDW To 100

[0216] Preparation: PL was dissolved in PG. In a separate vessel, KET was suspended in 20% of DDW, then Sodium Hydroxide Solution was added (to dissolve KET) and followed by the addition of Hydrochloric Acid (for pH adjustment). The Magnesium solution. and the rest amount of DDW were added. Finally, the PL solution was added to this solution and mixed with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany). Final pH-5.7

Example 16

Assessment of Phospholipid Magnesome Softness (More Fluid Bilayers Relative to Liposome) by Transition Temperature (Tm) Measurement by Differential Scanning Calorimetry (DSC)

[0217] Tm of the phospholipid was measured in the following compositions:

TABLE-US-00017 phospholipid Liposome Ingredients magnesome % w/w % w/w Magnesium sulfate 0.03 — PL 5 5 PG 15 — DDW To 100 To 100

[0218] The measurements were carried out using a Mettler Toledo DSC-1 STAR system (Toledo, China). Samples of 20 mg were placed in aluminum metal dishes. Tomograms were generated recording Tm values at a heating rate of 10° C./min within the temperature range of −50° C. to +50° C.

[0219] Results indicate that phospholipid magnesome systems had a Tm value of −7° C. vs. +7° C. for liposome. This lower Tm by 14° C. could be the result of a fluidization of the PL lamellae in phospholipid magnesome vesicles in comparison with classic liposome. The thermograms are shown in FIGS. 9A and 9B.

Example 17

Assessment of Drug Concentration in Plasma and Pharmacokinetic Parameters

[0220] Plasma concentration of Ketoprofen was measured following in vivo nasal administration and compared to oral administration. The experiment was carried out using Male Sprague Dawley (SD/Hsd) rats (Harlan, Israel).

[0221] The method of Ketoprofen extraction from plasma was validated according to FDA regulations for bio-analytical method validation, assessing precision, recovery, selectivity and linearity. The precision was 8.7%, the recovery was 96.6±5.5%, the limit of detection (LOD) was 0.84 mcg/ml, and the limit of quantification (LOQ) was 2.53 mcg/ml, Linear regression analysis of the plasma standard curve showed correlation with R.sup.2=1.00 over the concentration range of 0.05-100 mcg/ml Ketoprofen. The calibration curve equation: Y=143453X, (Y=Area, X=Concentration).

[0222] Ketoprofen (14 mg/kg) was administered nasally from Ketoprofen new nanovesicular carrier (KET-phospholipid Magnesome) and compared to oral administration (KET-PO). The compositions are tabulated below.

TABLE-US-00018 Compositions KET-phospholipid magnesome KET-PO Ingredients % w/w % w/w Ketoprofen (KET) 20 2 PL 5 — PG 30 — Sodium Hydroxide (10%) 40 40 Hydrochloric Acid (32%) 5.8 5.8 Magnesium sulfate 0.02 — Methyl Cellulose — 2 DDW To 100 To 100

[0223] Preparation: PL is dissolved in PG. In a separate vessel, KET is suspended in a part of water, then Sodium Hydroxide Solution is added (to dissolve KET) and followed by the addition of hydrochloric acid (for pH adjustment). The magnesium solution and the rest amount of DDW are added. Finally, the PL solution is added to this aqueous solution and mixed with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany) or Polytron homogenizer. Final pH-5.4

[0224] Blood samples were collected from rats' tails at 10, 30, 60, 120, 180, 240 and 300 min post drug administration. The blood samples were centrifuged at 3 k rpm for 10 min at 25° C. (HERMLE Z 160 M), and then 150 mcl of the plasma was taken. Plasma samples were frozen and kept at −20° C. until analysis.

[0225] A volume of 300 mcl of ACN was added to the plasma and mixed by vortex for 3 minutes at level 10, followed by the addition of 300 mcl of acetate buffer 0.05M with pH 5, and mixed by vortex for additional 1 minute at level 10. Then, the samples were centrifuged for 5 min at 14 k rpm at 25° C. (HERMLE Z 160 M), and the supernatants were filtered through Bulk GHP Acrodisc® 13 mm syringe filter with 0.45 um GHP membrane (Pall Corporation, USA), and transferred into pre-labeled auto injector vials before being injected into HPLC-UV.

[0226] The relative bioavailability (F %) was calculated according to the following equation:

F %=[(AUC.sub.NVC*DOSE.sub.PO)]/[(AUC.sub.PO*DOSE.sub.NCV)]*100

[0227] The AUC.sub.NVC and AUC.sub.PO represent the means of individual AUC from nasal and oral experimental groups, respectively.

[0228] The pharmacokinetic study aimed to evaluate the influence of the new nanovesicular carrier on the absorption parameters of the drug model, Ketoprofen. For this purpose, Ketoprofen concentration was assessed in plasma of rats following drug administration from new nanovesicular carrier as compared to oral administration. Ketoprofen plasma concentration curves versus time are plotted in FIG. 10 (rectangular: nasal administration according to the invention; triangle: oral administration.

[0229] Ketoprofen was assayed in rat plasma starting from 10 min post nasal or oral administration. Results are tabulated below.

TABLE-US-00019 PK parameters Nasal composition Oral administration T.sub.1/2,min 138.2 ± 21.1 328.7 ± 83.1   Tmax ,min 10.0 ± 0.0 94.0 ± 12.4  Cmax, mcg/ml 43.7 ± 8.3 13.7 ± 5.9   AUC.sub.0-5 hr 4130.4 ± 730.6 2642.1 ± 1153. 3 T.sub.last,min 300.00 ± 0.00  300 ± 0.00 Bioavailability 156.3 (relative to oral, %)

[0230] Results presented above show that very significant higher plasma concentrations (P<0.01) were detected 10 and 30 min after Ketoprofen administration in nasal vesicular nanocarrier. C.sub.max plasma values calculated for new nanovesicular carrier and oral administration were 43.65±8.30 and 13.68±5.91 mcg/ml, respectively. The T.sub.max values were 10 and 94 min for nasal vesicular carrier administration and oral administration, respectively.

[0231] These results above indicate that Ketoprofen was rapidly delivered from the nasal cavity to the systemic circulation following nasal administration showing that the phospholipid magnesome possesses enhanced delivery properties and enhanced effect for the first 90 minutes relative to the oral administration, with a very rapid onset of action and behaves similar to the oral administration for the next five tested hours.

Example 18

Cannabidiol-Containing Phospholipid Magnesome

[0232]

TABLE-US-00020 Ingredients % w/w CBD 5 PL 5 PG 30 Ethanol 15 Hemp Seed Oil (HSO) 4 Magnesium Sulfate 0.03 DDW To 100

[0233] PL is dissolved in PG and Ethanol mixture, and then HSO is added. CBD is dissolved in this solution. Then, Magnesium Sulfate aqueous solution (10 mg/ml) is slowly added through vigorous mixing with a homogenizer. Finally, DDW is slowly added with mixing.

Example 19

Cannabidiol-Containing Phospholipid Magnesome

[0234]

TABLE-US-00021 Ingredients % w/w THC 0.5 PL 5 VitE 0.5 PG 30 Olive Oil 3 Magnesium Sulfate 0.03 DDW To 100

[0235] PL is dissolved in PG, then Vit E, Oil and THC are added to the PL solution. To this solution, Magnesium Sulfate aqueous solution (10 mg/ml) is added slowly through mixing with an overhead stirrer (Heidolph digital 200 RZR-2000, Germany).

Examples 20-21

Insulin-Containing Phospholipid Magnesome

[0236]

TABLE-US-00022 Example 20 Example 21 ingredients ingredients % w/w % w/w Insulin 0.1 0.1 PL 3.0 3.0 PG 15.0 15.0 Vitamin E 0.3 0.3 Magnesium 1.0 5.0 sulfate DDW 80.6 76.6

[0237] Preparation: PL was dissolved in PG, then Vitamin E was added. In a separate vessel, Magnesium Sulfate was dissolved in DDW. The Magnesium solution was added gradually to the PL solution and mixed well. Finally, Insulin (from Bovine Pancreas—Sigma Aldrich, USA) was added and mixed well. The mixing through the entire preparation process was performed using an overhead Heidolph® stirrer (Heidolph Digital 200 RZR-2000, Germany).

Examples 22-23

Bacitracin-Containing Phospholipid Magnesome

[0238]

TABLE-US-00023 Example 22 Example 23 ingredients ingredients % w/w % w/w Bacitracin 0.1 0.1 PL 3.0 3.0 PG 15.0 15.0 Vitamin E 0.3 0.3 Magnesium sulfate 10 20 DDW 71.6 61.6

[0239] PL was dissolved in PG, then Vitamin E was added. In a separate vessel, Magnesium Sulfate was dissolved in DDW. The Magnesium solution was added gradually to the PL solution and mixed well. Finally, Bacitracin (Sigma Aldrich, USA) was added and mixed well. The mixing through the entire preparation process was performed using an overhead Heidolph® stirrer (Heidolph Digital 200 RZR-2000, Germany).

Example 24

Tramadol HCl—Containing Phospholipid Magnesome

[0240]

TABLE-US-00024 ingredients % w/w Tramadol HCl 1.0 PL 3.0 PG 15.0 Vitamin E 0.3 Magnesium sulfate 5.0 DDW 75.7

[0241] PL was dissolved in PG, then Vitamin E was added. In a separate vessel, Tramadol HCl (Chemagis, Israel) was dissolved in DDW and followed by dissolving the Magnesium Sulfate. The aqueous solution of Tramadol HCl and Magnesium was added gradually to the PL solution and mixed well. The mixing through the entire preparation process was performed using an overhead Heidolph® stirrer (Heidolph Digital 200 RZR-2000, Germany).

Example 25

Safinamide Mesylate—Containing Phospholipid Magnesome

[0242]

TABLE-US-00025 ingredients % w/w Safinamide mesylate 20 PL 5.0 PG 15.0 Vitamin E 0.5 Magnesium sulfate 0.5 DDW to 100

[0243] PL is dissolved in PG, then Vitamin E is added. In a separate vessel, Safinamide mesylate is dissolved in DDW and is added gradually to the PL solution and mixed well using an overhead Heidolph® stirrer (Heidolph Digital 200 RZR-2000, Germany). Finally, the Magnesium Sulfate is added to the composition and mixed.

Examples 26-34

Phospholipid Magnesome with Varying Mg Content

[0244]

TABLE-US-00026 Ex. 26 Ex. 27 Ex. 28 Ex. 29 Ex. 30 Ex. 31 Ex. 32 Ex. 33 Ex. 34 ingredients % w/w % w/w % w/w % w/w % w/w % w/w % w/w % w/w % w/w PL 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 PG 15.0 15.0 15.0 15.0 15.0 15.0 15.0 15.0 15.0 Vitamin E 0.3 0.3 0.3 0.3 0.3 0.3 0.2 0.3 0.3 Magnesium 0.01 0.1 0.3 0.5 1.0 2.0 5.0 10.0 20.0 sulfate DDW 81.69 81.6 81.4 81.2 80.7 79.7 76.7 71.7 61.7

[0245] PL was dissolved in PG, then Vitamin E was added. In a separate vessel, Magnesium Sulfate was dissolved in DDW. The Magnesium solution was added gradually to the PL solution and mixed well using an overhead Heidolph® stirrer (Heidolph Digital 200 RZR-2000, Germany).

Example 35

Pramipexole—Containing Phospholipid Magnesome

[0246]

TABLE-US-00027 ingredients % w/w Pramipexole 1.00 Propylene glycol (PG) 20.0 Phospholipon 90 G (PL) 3.00 α-D-tocopheryl acetate (Vit E) 0.50 Magnesium sulfate anhydrous (MgSO.sub.4) 0.10 Sodium hydroxide (NaOH) 0.05 Double distilled water (DDW) To 100

[0247] PL was mixed with PG using an overhead stirrer at 700 rpm (Heidolph, Hei Torque 200) until completely dissolved. Vit E was added and mixed well. MgSO.sub.4 was dissolved in about one third of the water amount, and the solution was added to the above PG solution through mixing.

[0248] In a separate vessel, NaOH 1% w/v solution was added to two thirds of the water amount. Pramipexole was dissolved in the above water solution. pH was measured and, if needed, adjusted to ˜4.5 with NaOH. This Pramipexole solution was then added through mixing at 700 rpm to the above system. The mixing was further continued for 5 min.

Example 36

Pramipexole—Containing Phospholipid Magnesome

[0249]

TABLE-US-00028 ingredients % w/w Pramipexole 0.50 Propylene glycol (PG) 20.0 Phospholipon 90 G (PL) 3.00 α-D-tocopheryl acetate (Vit E) 0.50 Magnesium sulfate anhydrous (MgSO.sub.4) 0.10 Sodium hydroxide (NaOH) 0.023 Double distilled water (DDW) To 100

[0250] PL was mixed with PG using an overhead stirrer at 700 rpm (Heidolph, Hei Torque 200) until completely dissolved. Vit E was added and mixed well. MgSO.sub.4 was dissolved in about one third of the water amount, and the solution was added to the above PG solution through mixing.

[0251] In a separate vessel, NaOH 1% w/v solution was added to two thirds of the water amount. Pramipexole was dissolved in the above water solution. pH was measured and, if needed, adjusted to ˜4.5 with NaOH. This Pramipexole solution was then added through mixing at 700 rpm to the above system. The mixing was further continued for 5 min.

Example 37

Effect of Nasal Administration of Pramipexole Phospholipid Magnesome Versus Drug Oral Administration in Mice Model for Parkinson Disease with Locomotor Impairment

[0252] The goal of the experiment reported below was to evaluate the effect of nasal administration of Pramipexole Phospholipid Magnesome on impaired locomotor activity in model mice for Parkinson's disease in comparison with oral administration of the drug and untreated animals. The animal model was obtained by administering Reserpine to mice.

[0253] Experimental Protocol

[0254] Compositions

[0255] The compositions tested were the one illustrated in Example 36 (0.5% w/w Pramipexole in Phospholipid Magnesome) and an aqueous solution of 0.5% w/w Pramipexole in water for oral administration, prepared by adding NaOH 1% w/v solution to DDW to achieve NaOH at concentration of 0.022% w/w, followed by dissolution of the drag in the alkaline solution.

[0256] Animals

[0257] All procedures carried out on animals were according to The National Institutes of Health regulations and were approved by the Committee for Animal Care and Experimental Use of the Hebrew University of Jerusalem.

[0258] The experiment was performed on fifteen male CD-1 ICR mice (27-32 g). Mice were housed under standard conditions of light and temperature in plastic cages in the specific-pathogen unit (SPF) of the pharmacy school at the Hebrew University of Jerusalem. Animals were kept in separated cages with smooth flat floor and provided with unlimited access to water and food.

[0259] Treatments

[0260] The mice were divided randomly into two drug treated groups, Pramipexole Phospholipid Magnesome administrated nasally (n=6), Pramipexole oral solution (n=4) and one untreated control group (n=5). Animals in the treatment groups received Pramipexole nasally from Phospholipid Magnesome or orally from solution at a dose of 3 mg/kg. Twenty minutes after the treatments, the behavioral testing was assessed. To rule out the effect of anesthesia, animals in the untreated control groups were anesthetized at the same time points before the behavioral testing.

[0261] On the first and eighth days of the experiment, the animals in the three groups received intraperitoneal injections of Reserpine at doses of 4 and 3 mg/kg, respectively. Reserpine injection was prepared in DDW containing 0.1% DMSO and 0.3% Tween 80. The suspension was further processed for 20 min at 50% power ratio using an Ultrasonic processor, Sonic-650WT-V2 Ultrasonic processor. Sonic Series, MRC Ltd, Holon, Israel.

[0262] Behavioral Testing

[0263] The behavioral tests (open field test and ptosis score) were performed on day 9 of the experiment, 23 hours after last Reserpine injection and 20 min following nasal or oral Pramipexole administration.

[0264] Open Field Test

[0265] Spontaneous locomotor activity of animals was measured using the open field test. Mice were placed in the center of a cage (29×28.5×30 cm), with the floor divided into nine equal squares. The number of squares crossed was counted during 5 min with no habituation session.

[0266] Normal animal moves in the cage and crosses squares on the floor. Reserpinized animal suffers from akinesia (cannot move) and crosses much less squares. Efficient treatment will reverse animal's behavior to normal.

[0267] Ptosis Score

[0268] Ptosis is the eye closure due to drooping of the upper eyelid. Reserpine induced ptosis was visually determined. Ptosis was recorded on a 0-4 scale, in which 0 represents eyes completely shut, and 4 completely open.

[0269] Normal animal has a ptosis score of 4. The score for reserpinized animals is reduced to 0-1. Efficient treatment will reverse the score to normal.

[0270] Results of the Open Field Test

[0271] The number of squares crossed (Mean±SD) in the open field test by Parkinson's mice model treated are tabulated below.

TABLE-US-00029 Nasal Pramipexole Phospholipid Pramipexole Group Magnesome Oral Untreated Number of 81.8 ± 25.3 27.5 ± 30.1 2.6 ± 3.6 squares crossed *Normal animals cross more than 100 squares.

[0272] The results are also presented graphically in the form of a bar diagram in FIG. 11.

[0273] The results pertaining to the open field test indicate that mice treated nasally with Pramipexole Phospholipid Magnesome expressed higher locomotor activity and crossed 81.8±25.3 squares. The animals in the Pramipexole orally treated and the untreated groups crossed only 27.5±30.1 and 2.6±3.6 squares, respectively (FIG. 11). The results show that the nasal administration of Pramipexole Phospholipid Magnesome significantly (p<0.01) enhanced the locomotor activity by 300% in comparison with oral treatment.

[0274] Results of Ptosis Score

[0275] Ptosis scores for Parkinson's mice model treated (Mean±SD) are tabulated below.

TABLE-US-00030 Nasal Pramipexole Phospholipid Pramipexole Group Magnesome oral Untreated Ptosis score 3.3 ± 0.7 2.0 ± 1.4 1.2 ± 0.4 *Normal animal has a ptosis score of 4.

[0276] The results are also presented graphically in the form of a bar diagram in FIG. 12.

[0277] In the evaluation of Reserpine-induced ptosis, a score of 3.3±0.7 was recorded for nasal Pramipexole Phospholipid Magnesome as compared to only 2.0±1.4 and 1.2±0.4 for the orally treated and the untreated groups, respectively (FIG. 12).

[0278] These results indicate enhanced anti-Parkinson's effect of Pramipexole achieved by means of nasal administration using Phospholipid Magnesome in comparison with oral administration.

Example 38

Effect of Nasal Administration of Pramipexole Phospholipid Magnesome Versus Drug Oral Administration in Mice Model for Parkinson Disease with Locomotor Impairment

[0279] The goal of the experiment was to evaluate the effect of nasal administration of Pramipexole Phospholipid Magnesome on impaired locomotor activity in model mice for Parkinson's disease in comparison with oral administration of the drug and untreated animals. The animal model was obtained by administrating Reserpine to mice.

[0280] Experimental Protocol

[0281] Compositions

[0282] The compositions tested were the one illustrated in Example 36 (0.5% w/w Pramipexole in Phospholipid Magnesome) and an aqueous solution of 0.5% w/w Pramipexole in water for oral administration, prepared by adding NaOH 1% w/v solution to DDW to achieve NaOH at concentration of 0.022% w/w, followed by dissolution of the drag in the alkaline solution.

[0283] Animals

[0284] All procedures carried out on animals were according to The National Institutes of Health regulations and were approved by the Committee for Animal Care and Experimental Use of the Hebrew University of Jerusalem.

[0285] This experiment was performed on 21 male CD-1 ICR mice (25-29 g). Mice were housed under standard conditions of light and temperature in plastic cages in the specific-pathogen unit (SPF) of the pharmacy school at the Hebrew University of Jerusalem. Animals were kept in separated cages with smooth flat floor and provided with unlimited access to water and food.

[0286] Treatments

[0287] The mice were divided randomly into two drug treated groups (Pramipexole Phospholipid Magnesome administrated nasally, and pramipexole oral solution) and one untreated control group (n=7/group). Animals in the treatment groups received Pramipexole at a dose of 3 mg/kg nasally from Phospholipid Magnesome or Pramipexole orally from solution, 20 min before the behavioral testing. To rule out the effect of anesthesia, animals in the untreated control groups were anesthetized at the same time points before the behavioral testing.

[0288] On the first and sixth days of the experiment, the animals of the three groups received an intraperitoneal injections of Reserpine at a dose of 3 mg/kg.

[0289] Behavioral Testing

[0290] The behavioral tests (open filed test, ptosis score and bar catalepsy test) were performed on day 7 of the experiment, 23 hours after last Reserpine injection. Open field test and ptosis score were assessed 20 min following nasal or oral Pramipexole administration. Bar catalepsy test was carried out immediately after the open field test (25 min following Pramipexole administration).

[0291] Open Field Test

[0292] The same protocol as in Example 37.

[0293] Ptosis Score

[0294] The same protocol as in Example 37.

[0295] Bar Catalepsy Test

[0296] Cataleptic immobility is regarded as an animal equivalent of akinesia and is demonstrated by an animal allowing its body to be placed in and maintain abnormal or unusual postures. We used the bar test to determine the ability of nasal administration of Pramipexole in Phospholipid Magnesome to reduce the duration of catalepsy in Parkinson's mice model in comparison with oral administration of the drug.

[0297] Twenty-five minutes after nasal and oral treatments, both fore paws of the mice were placed on a horizontal bar (diameter, 0.7 cm) 5 cm above the surface and then gently released. The catalepsy duration retained in this unusual position was recorded in seconds from the moment when an animal was released to the moment when it shifted its front paws from the initial position on the bar. The trial ended either when the animal started to move or after 60 s of immobility (cut off time).

[0298] Normal animal releases the bar immediately. Reserpinized animal suffers from catalepsy cannot return to normal position and therefore stays on the bar for longer time. Efficient treatment will reverse animal's behavior to normal.

[0299] Results of the Open Field Test

[0300] The number of squares crossed (Mean±SD) in the open field test by Parkinson's mice model treated are tabulated below.

TABLE-US-00031 Nasal Pramipexole Phospholipid Pramipexole Group Magnesome oral Untreated Number of 109.0 ± 29.8 37.4 ± 22.5 28.5 ± 32.4 squares crossed *Normal animals cross more than 100 squares.

[0301] The results are also presented graphically in the form of a bar diagram in FIG. 13.

[0302] The results pertaining to the open field test indicate that mice treated nasally with Pramipexole Phospholipid Magnesome expressed higher locomotor activity and crossed 109.0±29.8 squares. The animals in the Pramipexole orally treated and the untreated groups crossed only 37.4±22.5 and 28.5±32.4 squares, respectively (FIG. 13). The results are significant: p<0.01 for Pramipexole Phospholipid Magnesome versus Pramipexole oral and p<0.001 for Pramipexole Phospholipid Magnesome versus untreated control.

[0303] Results of the Bar Catalepsy Test

[0304] Catalepsy duration by the bar test (Mean±SD) for Parkinson's mice model treated are tabulated below.

TABLE-US-00032 Nasal Pramipexole Phospholipid Pramipexole Group Magnesome oral Untreated Catalepsy 0.8 ± 0.6 9.0 ± 8.3 21.3 ± 18.7 duration (sec) *catalepsy duration measured by bar test for normal animals is 0 sec.

[0305] The results are also presented graphically in the form of a bar diagram in FIG. 14.

[0306] The results pertaining to the bar catalepsy test indicate that nasal administration of Pramipexole from Phospholipid Magnesome reduced the catalepsy duration after 25 min from 21.3±18.7 in the reserpinized untreated animals to 0.8±0.6 sec (p<0.001). Oral administration of the same dose led to mild and non-significant reduction in the catalepsy duration (9.0±8.3 sec).

[0307] Results of Ptosis Score

[0308] Ptosis scores for Parkinson's mice model treated (Mean±SD) are tabulated below.

TABLE-US-00033 Nasal Pramipexole Phospholipid Pramipexole Group Magnesome oral Untreated Ptosis score 3.7 ± 0.5 1.7 ± 1.1 1.4 ± 1.0 *Normal animal has a ptosis score of 4.

[0309] The results are also presented graphically in the form of a bar diagram in FIG. 15.

[0310] In the evaluation of Reserpine induced ptosis, a score of 3.7±0.5 was recorded for Pramipexole Phospholipid Magnesome as compared to only 1.7±1.1 and 1.4±1.0 for the orally treated and the untreated groups, respectively (FIG. 15). The results of these experiments confirm the finding illustrated in Example 37 and show that the nasal administration of Pramipexole Phospholipid Magnesome significantly increased the locomotor activity by 300% in comparison with oral treatment. Furthermore, the nasal treatment completely reversed the reserpine induced catalepsy. These finding point towards the ability of nasal administration of Pramipexole Phospholipid magnesome to improve the treatment of Reserpine induced Parkinson's in mice.