N-palmitoyl ethanolamide for use in the prevention of colorectal cancer

Abstract

To provide use of N-palmitoyl-ethanolamide in antitumor therapy against colorectal carcinoma (CRC).SOLUTION: Described herein is a use of N-palmitoylethanolamide in the context of an antitumor therapy, in particular, a use of Palmitoylethanolamide in the prevention of colorectal carcinoma, adenomatous polyps, and / or pre-neoplastic ACF lesions. Also described herein are palmitoylethanolamide for use in the antineoplastic therapy of colorectal carcinoma, in which the palmitoylethanolamide exerts a selective antiproliferative action on tumor cells and not on healthy cells. The palmitoylethanolamide is in an ultra-micronized form having a particle size distribution, defined as percentage by volume and measured by the laser light scattering method, represented by a distribution curve having the mode below 6 microns and above 0.5 microns.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to the use of N-palmitoylethanolamide in anti-tumor therapy.

Background Art

[0002] Colorectal cancer (CRC) is a very common oncological disease and a major cause of death with a rapidly increasing incidence worldwide. In the United States, 147,950 people received a new diagnosis of CRC in 2020, of which 17,930 (12%) were estimated to be under 50 years old. In Europe, CRC was the second leading cause of cancer death in 2018.

[0003] In Italy, it is estimated that approximately 53,000 people are newly diagnosed with colorectal cancer (CRC) every year, and this disease ranks second in frequency among both men and women. Since the mid-2000s, the incidence of CRC has been steadily increasing due to an increase in risk factors that support the development of CRC.

[0004] Many factors, including lifestyle, unbalanced diet, genetic factors, changes in gut microbiota, inflammatory bowel disease (IBD), and other diseases such as Crohn's disease, ulcerative colitis, familial adenomatous polyposis (FAP), and Lynch syndrome, trigger the pathogenesis mechanism of this tumor.

[0005] The incidence of CRC began to decrease simultaneously with the introduction of a drug-prevention or chemo-prevention program consisting of the use of substances shown to reduce the risk of screening and the appearance of solid tumors.

[0006] Chemoprevention is a branch of oncology that targets healthy individuals at high risk of developing cancer and seeks to act within a broad timeframe during which molecular transformation processes lead to cancer. Therefore, preventive strategies aim to eliminate or reduce exposure to carcinogens, identify at-risk subjects, and intervene pharmacologically and / or dietarily to interrupt the carcinogenic process. Chemoprevention attempts to disrupt neoplastic transformation by acting on both the activation and proliferation phases.

[0007] The difference between chemotherapy and prophylactic treatment is significant, and the two approaches should not be confused: prophylactic treatment involves the use of substances that do not harm healthy cells and therefore the health of the person being treated. Conversely, chemoprevention involves the use of harmful chemicals directed not only at diseased cells but also at healthy cells. Thus, chemoprevention aims to create and maintain a certain homeostatic balance in order to prevent uncontrolled cell regeneration.

[0008] In humans, CRCs develop in the final part of the gastrointestinal tract and are caused by the malignant transformation of polyps following uncontrolled proliferation of intestinal mucosal cells. Although polyps are benign lesions, they are considered a precancerous form.

[0009] In most cases, CRCs begin as benign adenomatous polyps, from which highly malformed adenomas develop, and then progress to invasive cancer through the accumulation of genetic and epigenetic changes that determine the gradual loss of genetic stability.

[0010] The transformation process of normal to cancerous intestinal mucosal cells in the colorectal region arises from mutations in tumor suppressor genes, which are used to block cell cycle progression and protect cells from the accumulation of mutations that can lead to cancer. Without such blockages, cells progress towards transformation into cancer cells in an uncontrolled manner.

[0011] To date, the treatment aimed at eradicating CRC has been surgery, and increasingly, conservative interventions are being performed, often preceded by chemotherapy and radiotherapy to avoid so-called ostomies, if possible. The most active anticancer agents in the treatment of CRC include fluoropyrimidines (intravenous 5-fluorouracil, oral capecitabine), oxaliplatin, and irinotecan.

[0012] What I wanted to say above, particularly regarding the development of colorectal cancer, is therefore clear that there is a need to provide treatments that can prevent the development of such tumor forms, and, in the case of subjects already suffering from such a disease, provide safe treatments, that is, treatments that selectively act only on cancer cells and do not affect healthy cells. [Overview of the project]

[0013] This invention stems from the remarkable discovery that palmitoylethanolamide (PEA), when used preferably in an ultrafine powder form, can exert a preventive effect against the formation of colorectal cancer (CRC) and benign adenomatous polyps, as well as a specific antitumor effect against tumor cells.

[0014] The present invention relates to palmitoylethanolamide for use in the prevention of colorectal cancer, adenomatous polyps and / or preneoplastic ACF lesions.

[0015] The present invention also relates to palmitoylethanolamide for use in antitumor therapy for colorectal cancer, wherein palmitoylethanolamide exerts a selective antiproliferative effect against tumor cells rather than healthy cells.

[0016] Furthermore, the present invention includes the following aspects and embodiments. [1] Palmitoylethanolamide for use in the prophylaxis of colorectal cancer, adenomatous polyps and / or preneoplastic ACF lesions. [2] Palmitoylethanolamide for use in antitumor therapy for colorectal cancer, wherein palmitoylethanolamide exerts a selective antiproliferative effect against tumor cells rather than healthy cells. [3] Palmitoylethanolamide for use as described in [2], wherein PEA exerts a selective antiproliferative effect against tumor cells by arranging the cell cycle of tumor cells in the G2 / M phase. [4] Palmitoylethanolamide for use according to any of [1] to [3], wherein the palmitoylethanolamide is in an ultrafine form having a particle size distribution defined by volume percentage and measured by laser light scattering and represented by a distribution curve having modes less than 6 μm and greater than 0.5 μm. [5] Palmitoylethanolamide for use as described in [4], having a particle size distribution defined by volume percentage and measured by laser light scattering and measured in a Malvern Mastersizer 3000 instrument equipped with a Fraunhofer calculation algorithm, wherein at least 95 volume percent, preferably at least 99 volume percent, of the particles have a particle size of less than 6 μm. [6] Palmitoylethanolamide for use as described in [4], having a particle size distribution defined by volume percent and measured by laser light scattering method and measured on a Malvern Mastersizer 3000 instrument with Fraunhofer calculation algorithm, having a mode of 2-4 μm, with 100% by volume of particles smaller than 10 μm and at least 60% of particles smaller than 3 μm. [7] Palmitoylethanolamide for use as described in any of [1] to [6], wherein palmitoylethanolamide is administered once to four times a day at a dose of 10 mg to 1500 mg or 100 mg to 900 mg of PEA per dose unit. [8] Palmitoylethanolamide for use according to any of [1] to [7], comprising a pharmaceutical or veterinary preparation and formulated into dosage forms for oral, buccal, parenteral, rectal or transdermal administration. [9] Palmitoylethanolamide for use according to any of [1] to [7], wherein palmitoylethanolamide is included in a dietary composition, a nutritional supplement, or a food for special medical purposes (FSMP).

[10] Palmitoylethanolamide for use as described in any of [1] to [9], in which palmitoylethanolamide is used in chronic administration. These and further objects outlined in the appended claims are described in the following description. The body of the claims shall be considered to be included in the description for the purpose of assessing sufficiency of the description.

[0017] Further features and advantages of the present invention will become apparent from the following description of the preferred embodiments given as non-limiting examples.

Brief Description of the Drawings

[0018] [Figure 1] FIG. 1 shows a particle size distribution graph of palmitoylethanolamide in ultrafine form.

[0019] [Figure 2] FIGS. 2A, 2B, and 2C show graphs of the cell proliferation rates of HCT116 cells (A), Caco-2 cells (B), and healthy human colonic epithelial cells HCEC (C) for 24 hours in the presence or absence of ultrafine PEA, and the results are expressed as the cell proliferation percentage as mean ± SEM. *P < 0.05 and ***P < 0.001 compared to the control group, and evaluated by Student's t-test (ns = not significant).

[0020] [Figure 3]Figures 3A, 3B, and 3C show graphs of cell cycle arrest induction by ultrafine PEA. (A) Flow cytometry analysis of HCT116 cells in the presence or absence of ultrafine PEA (30 μM for 24 hours), with *P<0.05 and ****P<0.0001 compared to the control, as evaluated by Student's t-test. Gene expression of cyclin B1 (B) and cyclin-dependent kinase 1 (CDK1) (C) in HCT116 cells in the presence or absence of ultrafine PEA (30 μM for 24 hours). Gene expression was measured by real-time PCR, and the results are expressed as mean ± SEM. *P<0.05 and ***P<0.001 compared to the control group, as evaluated by Student's t-test.

[0021] [Figure 4] Figures 4A, 4B, and 4C show graphs of the effects of ultrafine PEA treatment in an AOM mouse model of colon cancer: (A) ACF, (B) polyps, and (C) total number of tumors derived from AOM administration (10 mg / kg at the start of weeks 1, 2, 3, and 4 of the experiment (corresponding to a total of 40 mg / kg by intraperitoneal administration)) in mice treated with or untreated with ultrafine PEA 10 mg / kg, administered orally three times a week for a 13-week experimental period, starting the week before the first AOM administration. Results are expressed as mean ± SEM. Compared to the control group treated with AOM alone, *P<0.05 and **P<0.01 were observed using Student's t-test. [Modes for carrying out the invention]

[0022] In a first embodiment, the present invention relates to palmitoylethanolamide (PEA) for use in the prevention of colorectal cancer (CRC).

[0023] The term "prevention" refers to pharmacological treatment of healthy patients, preferably those at risk of colorectal cancer, to prevent the development of neoplastic changes or neoplastic transformations of benign formations by acting on both the activation and proliferation stages.

[0024] In particular, the present invention allows for the detection of tumor and inflammatory biomarkers, and patients at risk of CRC who are diagnosed as positive for such biomarkers can be subjected to chronic PEA therapy, especially ultrafine PEA, to restore a proper gut environment to counteract and / or delay the onset of CRC.

[0025] In a second embodiment, the present invention relates to palmitoylethanolamide (PEA) for use in antitumor therapy of CRCs, wherein PEA exerts a selective antiproliferative effect against tumor cells rather than healthy cells.

[0026] In particular, the present invention relates to palmitoylethanolamide (PEA) for use in antitumor therapy of CRCs, wherein PEA exerts a selective antiproliferative effect on tumor cells by arranging the cell cycle of tumor cells in the G2 / M phase.

[0027] Preferably, the PEA is in an ultrafine form.

[0028] The term "ultrafine form of palmitoylethanolamide (or PEA)" means that the PEA has a particle size distribution defined by volume percentage and measured by laser light scattering, represented by a distribution curve having modes less than 6 μm and greater than 0.5 μm.

[0029] In one embodiment, the ultrafine form of PEA is measured using a Malvern Mastersizer 3000 instrument equipped with a Fraunhofer calculation algorithm, and has the particle size distribution defined above, wherein at least 95 volume percent, more preferably at least 99 volume percent, of the particles have a particle size of less than 6 μm.

[0030] In a particularly preferred embodiment, the ultrafine form of PEA is measured using a Malvern Mastersizer 3000 instrument equipped with a Fraunhofer calculation algorithm and has the particle size distribution defined above, with a mode of 2-4 μm, 100% by volume of particles smaller than 10 μm, and at least 60% of particles smaller than 3 μm.

[0031] Micronization can be carried out using a fluid jet system (e.g., a Jetmill® model system) operating with a helical technique using compressed air or nitrogen jets that can pulverize particles by utilizing kinetic energy instead of mechanical energy. Such devices are conventional and will therefore not be described further except as relating to the following features: - Inner diameter of the pulverization chamber: approximately 300 mm; - Fluid jet pressure 10-12 bar; - Product supply 9~12kg / h.

[0032] For the purposes of the present invention, PEA may be included in pharmaceutical or veterinary formulations and may be formulated into dosage forms for oral, buccal, parenteral, rectal, or transdermal administration.

[0033] For oral administration, pharmaceutical compositions may be in the form of tablets or hard or soft capsules and may be manufactured by conventional methods using pharmaceutically acceptable additives, such as binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or methylcellulose hydroxypropyl); excipients (e.g., lactose, crystalline cellulose, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or inhibitors (e.g., sodium lauryl sulfate). Tablets may be coated by methods well known in the art. Liquid formulations for oral administration may be in the form of liquids, syrups, or suspensions, or they may be lyophilized or granulated formulations that are reconstituted with water or other suitable vehicle before use. Such liquid formulations can be prepared by conventional methods using pharmaceutically acceptable additives, such as suspending agents (e.g., sorbitol syrup, cellulose derivatives, or edible hydrogenated fats); emulsifiers (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, or fractionated vegetable oils); and preservatives (e.g., methyl- or propyl-p-hydroxybenzoic acid or sorbic acid). The formulations may also conveniently contain flavorings, colorants, and sweeteners.

[0034] Formulations for oral administration can be appropriately formulated to allow for controlled release of the active ingredient.

[0035] For buccal administration, the composition may be in the form of tablets or pills formulated using conventional methods, suitable for absorption at the level of the buccal mucosa. A typical buccal formulation is a tablet for sublingual administration.

[0036] The compositions of the present invention can be formulated for non-enteral administration by injection. The injectable formulation may be provided as a single dose with added preservatives, for example, in a vial. The compositions may be provided in this form as a suspension, liquid, or emulsion in an oily or aqueous vehicle, and may contain formulation substances such as a suspension, stabilizer, and / or dispersant. Alternatively, the active ingredient may be in the form of a powder that is reconstituted with a suitable vehicle, such as sterile water, before use.

[0037] The compositions of the present invention can also be formulated according to rectal preparations such as suppositories or retained enemas, which may contain common suppository base ingredients such as cocoa butter or other glycerides.

[0038] In addition to the compositions described above, the compositions of the present invention may also be formulated as depot agents. Such long-acting formulations may be administered by implantation (e.g., subcutaneous, transdermal, or intramuscular) or intramuscular injection. Therefore, for example, the compositions may be formulated using a suitable polymer or hydrophobic material (e.g., in the form of an emulsion in a suitable oil) or an ion exchange resin, or as a minimally soluble derivative.

[0039] According to the present invention, the suggested dose of PEA for administration to a human (weighing approximately 70 kg) is in the range of 10 mg to 1500 mg or 100 mg to 900 mg of PEA per dose unit. The dose unit may be administered, for example, 1 to 4 times per day. The dose depends on the form in which the PEA is administered, i.e., whether it is non-micronized PEA, micronized PEA, or ultramicronized PEA. The dose also depends on the route of administration selected. It should be considered that the dose may need to be continuously adjusted depending on the patient's age and weight, as well as the severity of the clinical condition to be treated. The exact dose and route of administration are ultimately left to the discretion of the attending physician or veterinarian.

[0040] Regarding prophylactic treatment, PEA can be administered for long-term or chronic prophylactic treatment due to its extremely low toxicity.

[0041] The present invention further relates to dietary compositions, nutritional supplements, and foods for special medical purposes (FSMPs) comprising PEA, preferably ultrafine PEA, for use in preventing the development of CRC in healthy subjects.

[0042] The term "food for special medical purposes" refers to products approved in accordance with Regulation (EU) 2016 / 128. Such terms refer to products administered under medical supervision, and therefore equivalent to drugs.

[0043] The formulations according to the present invention can be manufactured according to conventional methods, such as those described in Remington's Pharmaceutical Sciences Handbook, Mack Pub. Co., NY, USA, 17th edition, 1985. [Examples]

[0044] (Miniaturization procedure)

[0045] The PEA was miniaturized as described above.

[0046] Ultra-miniaturization was achieved using a fluid jet system (particularly the Jetmill® model system) that operates with compressed air jet "helical technology".

[0047] Optimal miniaturization conditions: - Inner diameter of the pulverization chamber: 300 mm; - Fluid jet pressure 8 bar; - Product supply 9~12kg / h.

[0048] (Determination of particle size distribution)

[0049] The particle size distribution was determined using a wet sample after 1 minute of sonication.

[0050] The Malvern Mastersizer 3000 instrument, operating with LALLS (Low-Angle Laser Scattering) technology and Fraunhofer calculation algorithms, was used.

[0051] The particle size distribution graph is shown in Figure 1.

[0052] (Biological experiment)

[0053] In in vitro experiments, immortalized human colon cancer (HCT116), human colorectal adenocarcinoma cell lines (Caco-2), and immortalized healthy human colon epithelial cells (HCEC) were cultured in DMEM growth medium (Sigma Aldrich) supplemented with 10% fetal bovine serum (FBS, Sigma Aldrich) and maintained at 37°C in an incubator with a 5% CO2 atmosphere.

[0054] To assess viability, cells were stained with trypan blue, and cell proliferation was measured using BrdU proliferation ELISA kits (Roche, Milan, Italy) in or without ultrafine PEA (1-30 μM).

[0055] Cell cycle analysis of HCT116 cells, BD Pharmingen TM The study was conducted according to the protocol reported in the BrdU Flow Kit (BD Biosciences, USA). 5 Cells were seeded in 6-well plates and stored in serum-free medium for 24 hours in or without ultrafine PEA (30 μM). Cells were detected using a BriCyte flow cytometer (Mindray, Italy) according to their BrdU (bromodeoxyuridine) and 7-AAD (7-aminoactinomycin D) content. The obtained data were analyzed using FlowJo v10 software (tree Star, USA) to understand the distribution of HCT116 at various cell cycle stages.

[0056] The expression of the CDK1 and CYCLIN B1 genes was evaluated using quantitative real-time PCR with specific primers.

[0057] In vivo studies were conducted using 6-week-old (25-30g) male CD1 mice, which were allowed to feed freely and housed in cages with controlled sleep / wake cycles. Prior to the start of the experiment, the animals were subjected to a one-week acclimatization period, adhering to the principles of animal care approved by the Italian Ministry of Health, respecting the ARRIVE guidelines, and considering all experimental procedures and protocols (Curtis MJ et al. Experimental design and analysis and their reporting II: updated and simplified guidance for authors and peer reviewers. Br J Pharmacol, 2018; 175: 987-993).

[0058] Animals were chemically treated with AOM (azoxymethane, total 40 mg / kg) (Neufert C et al. An inducible mouse model of colon carcinogenesis for the analysis of sporadic and inflammation-driven tumor progression. Nat Protoc 2007; 2: 1998-2004) to induce CRC formation. A single dose of AOM at a concentration of 10 mg / kg was administered intraperitoneally at the start of weeks 1, 2, 3, and 4 of the experiment (Pagano E et al. Pharmacological inhibition of MAGL attenuates experimental colon carcinogenesis, Pharmacol Res 2017; 119: 227-236).

[0059] The animals were randomized into two groups, each consisting of 7 animals: Group 1 was treated orally with 2% carboxymethylcellulose (CMC), which is used as a vehicle for suspending ultrafine PEA. Group 2 was orally treated three times a week with ultrafine PEA in 2% CMC at a concentration of 10 mg / kg to evaluate chemopreventive effects, starting the week before the first dose of AOM (Pagano E et al. Pharmacological inhibition of MAGL attenuates experimental colon carcinogenesis, Pharmacol Res 2017; 119: 227-236).

[0060] The animals were euthanized 12 weeks after the initial AOM injection, and the colorectal region was collected.

[0061] Based on laboratory experience, the timing and dosage of AOM used ensured the development of a significant number of ACFs, polyps, and tumors (Izzo AA et al. Increased endocannabinoid levels reduce the development of precancerous lesions in the mouse colon. J Mol Med (Berl) 2008; 86: 89-98).

[0062] ACF, polyps, and colorectal tumors were detected and quantified according to the protocol described in Pagano E et al. Pharmacological inhibition of MAGL attenuates experimental colon carcinogenesis, Pharmacol Res 2017; 119: 227-236.

[0063] (result)

[0064] (In vitro proliferation of HCT116 and Caco-2 cells) Ultrafinely processed PEA significantly reduces the proliferation of HCT116 and Caco-2 cells in vitro; in particular, increased doses of ultrafinely processed PEA (1–30 μM) reduce the proliferation rate of HCT116 cells during 24-hour exposure (Figure 2A). Furthermore, 30 μM of ultrafinely processed PEA significantly reduces the proliferation of non-metastatic Caco-2 cells (Figure 2B). Surprisingly, the antiproliferative effect of ultrafinely processed PEA did not affect the proliferation rate of healthy epithelial cell lines HCEC (Figure 2C), demonstrating a clear selective effect against tumor cells. This data is extremely important as it allows for a significant reduction in side effects.

[0065] (Cell cycle arrest in G2 / M phase of HCT116 cells in vitro) Flow cytometry revealed that treatment of HCT116 cells with ultrafine PEA (30 μM for 24 hours) blocked G2 / M phase neoplastic cells and simultaneously significantly reduced the proportion of S phase cells (Figure 3A). Since G2 / M transition is known to be regulated by the cyclin B1 / CDK1 complex (Malumbres M and Barbacid M, 2005), we investigated the behavior of such a complex in the presence of ultrafine PEA and found that treatment of cells with ultrafine PEA increased the expression of the cyclin B1 / CDK1 complex (Figures 3B-C). These data indicate that ultrafine PEA can actively participate not only in the arrest of G2 / M phase neoplastic cells but also in the activation of the cyclin B1 / CDK1 complex, which is necessary for cell cycle arrest at the above stage.

[0066] (Prevention of tumor development in animal models of CRC) The administration of the carcinogen AOM (azoxymethane) is a risk factor for ACF. Abnormal foveal lesions, also known as (Figure 4A), polyps (Figure 4B), and tumors (Figure 4C) )of How many KanoIt induces formation. Treatment of animals with ultrafine PEA 10 mg / kg, administered orally three times a week for a 13-week experimental period, starting the week before the first AOM administration, significantly reduced the number of preneoplastic ACF lesions (Figure 4A) and the total number of tumors (Figure 4C) induced by the carcinogen AOM. Ultrafine PEA also showed a strong tendency to reduce the number of polyps (Figure 4B).

[0067] In light of these results, prophylactic administration of ultrafine PEA appears to be effective in reducing the incidence of CRC by delaying the transformation of intestinal mucosal cells into tumor cells (chemotherapy prophylactic effect). This data shows how prophylactic intake of ultrafine PEA is not only completely safe, but also significantly reduces the aggressiveness of CRC and effectively improves life expectancy.

[0068] Furthermore, the slow transformation of mucosal cells allows for treatment at the time of diagnosis of small CRCs, and possibly at an early stage when they are not yet aggressive.

[0069] The data obtained show for the first time how chemical preventive measures using PEA, particularly ultrafine PEA, can reduce the incidence of CRC.

[0070] Next, the present invention will be further explained using the following formulation examples.

[0071] (Example of formulation) PEA-UM = Ultrafine Palmitoyl Ethanolamide

[0072] Example 1 Each tablet contains: PEA-UM 300.00mg Crystalline cellulose 78.47 mg Croscarmellose sodium 45.00 mg Polyvinylpyrrolidone 10.00 mg Magnesium stearate 4.0 mg Polysorbate 80 2.00 mg

[0073] Example 2 Each tablet contains: PEA-UM 600.00mg Crystalline cellulose 156.94 mg Croscarmellose sodium 90.00 mg Polyvinylpyrrolidone 20.00 mg Magnesium stearate 8.00 mg Polysorbate 80 4.00 mg

[0074] Example 3 A 5g dose of orally disintegrating microgranules contains the following: PEA-UM 500.00mg Non-carious sugar 200.00mg Allowable additives up to 5.00g as needed.

[0075] Example 4 Hard gelatin capsules with appropriate coatings to make them stomach-resistant include the following: PEA-UM 400mg Soy lecithin 100mg Lactose 80mg

[0076] Example 5 Tablets coated with a stomach-resistant layer include: PEA-UM 500mg Micronized PEA 250mg Polyvinylpyrrolidone 30mg Croscarmellose sodium 80mg Magnesium stearate 7mg

[0077] Example 6 Suppositories include: PEA-UM 300mg Non-micronized PEA 500mg Add up to 3g of lipophilic base to the suppository as needed.

[0078] Example 7 2.5ml of oily gel contains: PEA-UM 600.00mg Glyceryl monostearate 40.00 mg Add up to 2.50 ml of vegetable oil as needed.

Claims

1. A composition comprising palmitoylethanolamide for use in the prevention of colorectal cancer, adenomatous colon polyps and / or preneoplastic ACF (abnormal foveal lesion) lesions, wherein the palmitoylethanolamide acts in both the activation and proliferation stages, and the composition is a pharmaceutical preparation, a veterinary preparation, a dietary composition, a nutritional supplement, or a food for special medical purposes (FSMP), and is formulated into a dosage form for oral administration.

2. The composition according to claim 1, wherein the palmitoylethanolamide is in an ultrafine form having a particle size distribution defined by volume percentage and measured by laser light scattering and measured with a Malvern Mastersizer 3000 instrument equipped with a Fraunhofer calculation algorithm, and represented by a distribution curve having modes less than 6 μm and greater than 0.5 μm.

3. The composition according to claim 2, having a particle size distribution defined by volume percentage and measured by laser light scattering method and measured with a Malvern Mastersizer 3000 instrument equipped with a Fraunhofer calculation algorithm, wherein at least 95 volume percent, preferably at least 99 volume percent, of the particles have a particle size of less than 6 μm.

4. The composition according to claim 2, wherein the palmitoylethanolamide has a particle size distribution defined by volume percentage and measured by laser light scattering and measured with a Malvern Mastersizer 3000 instrument equipped with a Fraunhofer calculation algorithm, having a mode of 2 to 4 μm, with 100% by volume of particles smaller than 10 μm and at least 60% by volume of particles smaller than 3 μm.

5. The composition according to any one of claims 1 to 4, wherein palmitoylethanolamide is administered once to four times a day at a dose of 10 mg to 1500 mg or 100 mg to 900 mg of palmitoylethanolamide per dose unit.

6. The composition according to any one of claims 1 to 5, wherein palmitoylethanolamide is used for chronic administration.

Images