WO2018162271A1

WO2018162271A1 - Nanoparticles for the treatment of macular degeneration

Info

Publication number: WO2018162271A1
Application number: PCT/EP2018/054710
Authority: WO
Inventors: Rita Dobmeyer
Original assignee: Rita Dobmeyer
Priority date: 2017-03-05
Filing date: 2018-02-26
Publication date: 2018-09-13

Abstract

The invention relates to a topical composition for the treatment or amelioration of macular degeneration (MD), comprising one or more active ingredients, which are efficient for the treatment of MD by intraocular injection, or a pharmaceutically acceptable salt thereof, wherein said active ingredient in a nanonized form.

Description

NANOP ARTICLES FOR THE TREATMENT OF MACULAR DEGENERATION

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit from the priorities of European patent applications EP 17159277.7 of March 5, 2017 and EP 17205967.7 of December 7, 2017; the entire of these applications is hereby incorporated by reference.

BACKGROUND OF THE INVENTION

1. TECHNICAL FIELD

The invention relates to a topical composition for the treatment or amelioration of macular degeneration (MD), comprising one or more active ingredients, which are efficient in the treatment of MD by intraocular injection, or a pharmaceutically acceptable salt thereof, wherein said active ingredient is in a nanonized form.

2. BACKGROUND INFORMATION

When HIV infects a cell, reverse transcriptase copies the viral single stranded RNA genome into a double-stranded viral DNA. The viral DNA is then integrated into the host chromosomal DNA, which then allows host cellular processes, such as transcription and translation, to reproduce the virus. NRTIs block reverse transcriptase's enzymatic function and prevent completion of synthesis of the double-stranded viral DNA, thus preventing HIV from multiplying.

Recently, it has been found that NRTIs possess intrinsic anti- inflammatory activity (Benjamin J. Fowler et al, Science. 2014 November 21; 346(6212): 1000-1003) and can be used to treat macular degeneration (W. Dunham Science News ,21 November 2014 "HIV drugs could treat blindness" in http://www.iol.co.za/scitech/science/news/hiv-drugs- could-treat-blindness- 1783926). Jayakrishna Ambati, M.D. published in Science, 2014 Nov 21; 346(6212): 1000-1003, that NRTIs block inflammation. In mice, the NRTIs stavudine and zidovudine prevented the progression of geographic atrophy, also known as dry MD. MD is a leading cause of blindness among people age 60+ worldwide, and approximately 90 percent of people with MD have geographic atrophy. There are currently no FDA-approved treatments for geographic atrophy.

MD involves deterioration of the retina, the light-sensitive tissue at the back of the eye. The most severe damage occurs in the macula, a small area of the retina that is needed for sharp, central vision. In geographic atrophy, there is a gradual breakdown of the nerve cells in the macula and the supporting cells beneath it, called retinal pigment epithelial (RPE) cells.

The protective effect of NRTIs is independent of their ability to prevent reverse

transcription. Instead, NRTIs block the inflammasome, a large protein complex that promotes inflammation. For this reason, it is suggested that NRTIs could have similar benefits for other inflammatory disorders, including neovascular MD, also called wet MD, a disease in which leaky blood vessels develop around the macula to cause blindness. Although several effective drugs for neovascular MD exist, they are given by regular injections into the eye and can become less effective with time.

Macular degeneration (MD) causes cells to die in the macula, a part of the eye located near the centre of the retina that permits vision in fine detail. Age related macular degeneration is the leading cause of vision loss of those over the age of 65 in the industrialized world. The prevalence and need to develop effective treatments for MD has led to the

development of multiple animal models. MD is a complex and heterogeneous disease that involves the interaction of both genetic and environmental factors. MD affects an estimated 50 million people worldwide. With the aging of the population, it is likely to affect 200 million people by the year 2020.

MD can for example be treated with laser coagulation, and more commonly with medication that stops and sometimes reverses the growth of blood vessels. Another treatment of wet MD is an intraocular shot of corticosteroids such as

dexamethasone or triamcinolone acetonide, or an anti-VEGF drug, which inhibits the formation of new blood vessels behind the retina and may keep the retina free of leakage, e.g. http://www.rvscny.com/intraocularinjections.html. An injection in the eye can be a disconcerting experience with serious side effects, and it may take several treatments to become accustomed to the procedure. The effect lasts for a month or maybe more.

The US patent US 6,217,895 teaches sustained release methods and devices for administering corticosteroids to a posterior segment of the eye. These devices are also inserted by intraocular injection.

The intraocular injections including the insertion of implanted devices are not well accepted by most of the patients. The problem underlying the present invention was to provide a composition for the treatment of MD, which avoids intraocular injections and can be administered locally to the eye in a low effective dose with reduced side effects.

It has been found surprisingly that active ingredients, which are efficient for the treatment of MD by intraocular injection, can be used to treat or ameliorate MD, if they are administered locally in a nanonized form.

BRIEF SUMMARY OF THE INVENTION Therefore, the invention relates to a topical composition for the treatment or amelioration of macular degeneration (MD), comprising one or more active ingredients, which are efficient for the treatment of MD by intraocular injection, or a pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable excipient, wherein said one or more active ingredients are in the form of nanoparticles. Furthermore, the invention relates to a nucleoside reverse transcriptase inhibitor (NRTI), a serine protease inhibitor (serpin), an immunosuppressant, a tetracycline or bile acid or a pharmaceutically acceptable salt or prodrug thereof for the treatment or amelioration of macular degeneration, wherein the NRTI, serpin or or an immunosuppressant is in nanonized form.

Another aspect of the invention is a method for the treatment of macular degeneration said method comprises administering an effective amount of one or more active ingredients, which are efficient for the treatment of MD by intraocular injection, or a pharmaceutically acceptable salt thereof in nanonized form to a patient in need thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGURE 1 shows the particle-size of different nanonized Al AT formulations over time. The formulations were stored up to 7 days in room temperature. Results are shown as ± SD (n+3).

FIGURE 2 shows the Count Rate of different nanonized Al AT formulations over time. The formulations were stored up to 7 days in room temperature. Results are shown as ± SD (n+3). DETAILED DESCRIPTION OF THE INVENTION

The term macular degeneration (MD) as used hereinbefore or hereinbelow includes age- related macular degeneration (AMD), both in the wet and the atrophic form, and diabetic macular degeneration (DMD).

The term "active ingredients, which are efficient for the treatment of MD by intraocular injection" as used hereinbefore or hereinbelow relates to those active principles that can be used to treat MD by intravitreal or subconjunctival injections into the eye, or via an implanted sustained release device delivering to the vitreous of a patient suffering from MD. These active ingredient embrace not only those, which have a proven efficiency against MD by intraocular injections, but also such active ingredients, of which the efficacy against MD by intraocular injection is highly likely in view of their mode of action.

These active ingredients include nucleoside reverse transcriptase inhibitors, serine protease inhibitors, immunosuppressants and statins.

The term "nucleoside reverse transcriptase inhibitor (NRTI)" as used hereinbefore or hereinbelow relates to small organic molecules that block reverse transcriptase's enzymatic function and prevent completion of synthesis of the double-stranded viral DNA, thus preventing HIV from multiplying. In order to be incorporated into the viral DNA, NRTIs must be activated in the cell by the addition of three phosphate groups to their deoxyribose moiety, to form NRTI triphosphates. This phosphorylation step is carried out by cellular kinase enzymes. Preferred are those NRTIs, which are approved by the US FDA and/or the EMA including Abacavir, Didanosine, Emtricitabine, Lamivudine, Stavudine, Zalcitabine and Zidovudine, most preferred are Stavudine and Zidovudine.

Stavudine (d4T), which is sold under the brand name Zerit®, is 2',3'-didehydro-2',3'- dideoxythymidine of formula

Zidovudine is also known as azidothymidine (AZT), l-[(2R,4S,5S)-4-azido-5- (hydroxymethyl)oxolan-2-yl]-5-methylpyrimidine-2,4-dione of formula

The term "serine protease inhibitor (serpin)" as used hereinbefore or hereinbelow relates to superfamily of related proteins that included both protease inhibitors (e.g. alpha 1- antitrypsin). The name "serpin" was coined based on the most common activity of the superfamily. Most serpins are protease inhibitors, targeting extracellular, chymotrypsin- like serine proteases. These proteases possess a nucleophilic serine residue in a catalytic triad in their active site. Examples include thrombin, trypsin, and human neutrophil elastase. Serpins act as irreversible, suicide inhibitors by trapping an intermediate of the protease's catalytic mechanism.

The most preferred serpin is Alpha- 1 Antitrypsin or ai -antitrypsin (Al AT), which is commercially available under the tradenames Prolastin® or Zemaira®. It is generally known as serum trypsin inhibitor. Alpha 1 -antitrypsin is also referred to as alpha- 1 proteinase inhibitor (A1PI) because it inhibits a wide variety of proteases. It protects tissues from enzymes of inflammatory cells, especially neutrophil elastase, and has a reference range in blood of 1.5 - 3.5 gram/liter.

The term "immunosuppressant" as used hereinbefore or hereinbelow relates to active ingredients that suppress the immune system and reduce the risk of rejection of foreign bodies such as transplant organs. Different classes of immunosuppressive agents have different mechanism of action. Now immunosuppressive agents are used as cancer chemotherapy, in autoimmune diseases such as rheumatoid arthritis and to treat severe allergy. As immunosuppressive agents lower the immunity there is increased risk of infection. Preferred immunosuppressants are azathioprine, lenalidomide, methotexate, omalizumab, pomalidomide and thalidomide and hydrocortisone derivatives including but not limited to prednisolone, prednisone, methylprednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide and loteprednol. Most preferred are prednisolone of formula,

fluocinolone acetonide of formula

triamcinolone acetonide of formula

dexamethasone of formula

and thalidomide (R,S)-a-(N-phthalimido)glutarimide of formula

The term "statins" as used hereinbefore or hereinbelow relates to 3-hydroxy-3- methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors. Preferred statins are atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin.

Most preferred is lovastatin of fo

The term "tetracyclines" as used hereinbefore or hereinbelow relates to a subclass of polyketides having an octahydrotetracene-2-carboxamide skeleton. Preferred tetracyclines are doxycycline of formula

and minocycline of formula

The term "bile acid" as used hereinbefore or hereinbelow relates to steroid acids found predominantly in the bile of mammals and other vertebrates. Bile acids are conjugated with taurine or glycine in the liver, and the sodium and potassium salts of these conjugated bile acids are called bile salts. Primary bile acids are those synthesized by the liver. Secondary bile acids result from bacterial actions in the colon.

Most preferred is tauroursodeox cholic acid (TUDCA) of formula,

which is an ambiphilic bile acid. It is the taurine conjugate form of ursodeoxycholic acid (UDCA). Humans are found to have only trace amounts of TUDCA. However, bears contain large amounts of TUDCA in their bile; UDCA and conjugates comprise about 47% of the bile in American black bears and up to 76% in Asiatic bears.

The term "nanonized" or "nanoparticle" as used hereinbefore or hereinbelow is intended to mean a material, which has been deliberately manipulated to have dimensions between 0.5 to 1000 nm, preferably having a mean particle size with an average diameter in the approximate range of 0.5 to 900 nm. Preferably, the nanoparticles will be in the range of 1 to 500 nm, more preferably in the range of 5 to 300 nm and most preferably 10 to 200 nm. In some instances, it is preferred that the nanoparticles are in an upper range of around 50 to 220 nm, more preferred in the range of 70 to 210 nm, even more preferred in the range of 75 to 200 nm and most preferred in the range of 80 to 190 nm. In other instances, it is preferred that the nanoparticles are in a lower range of 0.5 to 10 nm, more preferred in the range of 0.5 to 8 nm, even more preferred 1 to 7 nm and most preferred about 7 nm or lower. In another preferred embodiment the mean particles size of the nanoparticles is in the range of 1 nm to 100 nm, more preferred 20 to 90 nm. The dimensions indicated above relate to mean particles sizes, which are determined by dynamic light scattering (DSM) in aqueous environment, e.g. US 3,879,129 A or ISO 22412:2017 or atomic force microscopy (AFM) on a solid carrier, e.g. US 4,724,318 A or ASTM E2859-11(2017).

In a first embodiment the nanoparticles of the active ingredients are formed by specific polymers. It is preferred that the polymer comprises a linear and/or branched or cyclic polymonoguanide / polyguanidine, polybiguanide, analogue or derivative thereof. The formation of the nanonized complexes between the NRTIs, serpin,

immunosuppressants, statins, tetracyclines or bile acids and the linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide is carried out analogously as described in the US patent applications US 2014/242097 and US 2016/206567. In another embodiment the nanoparticles of the active ingredient are obtainable by a process, wherein herein a fluid dispersion containing the active ingredient is contacted with a supercritical fluid anti-solvent such as carbon dioxide under near-or supercritical temperature and pressure conditions for maximizing small particle formation. The formation of the nanonized NRTIs, serpin, immunosuppressants, statins, tetracyclines or bile acids can be carried out analogously as described the US patents US 5,833,891 and US 6,113,795.

In a preferred embodiment nanonized Al AT, AZT, prednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide, thalidomide, lovastatin, doxycycline or TUDCA is prepared in the presence of a nucleonic acid such as adenosine triphosphate (ATP), a rhamnolipid biosurfactant and/or chitosan.

Rhamnolipid biosurfactants are glyco lipids containing L-rhamnose and β-hydroxyl fatty acids, with amphiphilic properties (both hydrophilic and hydrophobic). The rhamnolipid products are obtained by purification from Pseudomonas aeruginosa and contain a mixture of rhamno lipids with fatty acids of varying tail length. They are highly biodegradable, nontoxic, and renewable. As a rule they reduce the surface tension of water to 25 to 40 mN/m, having a critical micelle concentration (CMC) of 5-380 mg/L, and decreasing the interfacial tension of oil and water to <1 dyne/cm. Most preferred is the rhamnolipid R90 (rhamno lipids, 90%), which is commercially available from Sigma- Aldrich, e.g.

http://www.sigmaaldrich.corn/catalog/product/sigma/r90?lang=de&region=DE.

Chitosan is a linear polysaccharide composed of randomly distributed P-(l→4)-linked D- glucosamine (deacetylated unit) and N-acetyl-D-glucosamine (acetylated unit). It may be useful in bandages to reduce bleeding and as an antibacterial agent; it can also be used to help deliver drugs through the skin.

The term "pharmaceutically acceptable salts" is meant to include salts of the compounds of the invention, which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric,

dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, salts of organic acids like glucuronic or galactunoric acids and the like and nucleosides such as adenosine triphosphate (ATP) (see, for example, Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

As used herein, the term "prodrug" relates to compounds which are quickly transformed in vivo into pharmacologically active compounds. The design of prodrugs is generally studied in Hardma et al. (Eds.), Goodman and Gilman's The Pharmacological Basis of

Therapeutics, 9th ed., pages 11-16 (1996). An in-depth study is carried out in Higuchi et al, Prodrugs as Novel Delivery Systems, Vol. 14, ASCD Symposium Series, and in Roche (ed.), Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987).

As used herein, the term "pharmaceutically acceptable" relates to molecular entities and compositions that are physiologically tolerable and do not normally cause an allergic reaction or a similar adverse reaction, such as gastric discomfort, dizziness and the like, when administered to humans. As used herein, the term "pharmaceutically acceptable" preferably means that it is approved by a regulatory agency of the federal or state government or listed in the US pharmacopoeia or another pharmacopoeia, generally recognized for its use in animals, preferably in mammals and more particularly in human beings.

By "effective" amount of a drug, formulation, or permeant is meant a sufficient amount of an NRTI, serpin, immunosuppressant or statin to provide the desired local or systemic effect. A "pharmaceutically effective" or "therapeutically effective" amount refers to the amount of NRTI or serpin needed to effect the desired therapeutic result.

Dosage levels of the order of from about 0.5 mg to about 100 mg per kilogram of body weight per day and more preferably from about 1.0 mg to about 50 mg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions. Dosage unit forms will generally contain between from about 1.0 mg to about 30 mg of the nanonized NRTI, nanonized immunosuppressant or nanonized statin per dose.

Dosage levels of the order of from about 0.5 μg to about 100 μg per kilogram of body weight per day and more preferably from about 1.0 μg to about 80 μg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions. Dosage unit forms will generally contain between from about 1.0 μg to about 50 μg of the nanonized serpin per dose. Frequency of dosage may also vary depending on the NRTI, serpin, immunosuppressant or statin used and the stage of the disease treated. However, a dosage regimen of 4 times daily or less is preferred. Most preferred is a once daily administration. It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration and rate of excretion, drug combination and the severity of the particular disease undergoing therapy.

Preferably, the ophthalmic formulation according to the administered 3 times a day with 10 to 20 drops.

The term "pharmaceutically acceptable excipient" or "pharmaceutically acceptable vehicle" refers to any formulation or excipient medium that provides the appropriate delivery of an effective amount of the active agents as defined herein, does not interfere with the effectiveness of the biological activity of the NRTI, the serpin,

immunosuppressant or statin, and that is sufficiently non-toxic to the host or patient.

Representative excipients include water, oils, both vegetable and mineral, cream bases, lotion bases, ointment bases and the like. These bases include suspending agents, thickeners, and the like. Their formulation is well known to those in the art of cosmetics and topical pharmaceuticals. Additional information concerning carriers can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005) which is incorporated herein by reference. "Pharmaceutically acceptable topical excipient" and equivalent terms refer to

pharmaceutically acceptable excipient, as described herein above, suitable for topical application. An inactive liquid or cream vehicle capable of suspending or dissolving the active agent(s), and having the properties of being nontoxic and non- inflammatory when applied to the eye, skin, nail, hair, claw or hoof is an example of a pharmaceutically acceptable topical carrier.

For ophthalmic use, the composition of the invention can be formulated using nanoparticle systems in isotonic, pH adjusted, sterile saline, or, preferably, as solutions in isotonic, pH adjusted, sterile saline, optionally in combination with a preservative such as a

benzylalkonium chloride.

Preferably, the ophthalmic composition comprises 1.0 wt. % to 5.0 wt. %, preferably 1.2 wt. % to 4.0 wt. %, in particular 1.5 wt. % to 3.0 wt. % of the NRTI, immunosuppressant statin, tetracycline or bile acid in nanonized form.

Preferably, the ophthalmic composition comprises 0.1 wt. % to 0.5 wt. %, preferably 0.12 wt. % to 0.4 wt. %, in particular 0.15 wt. % to 0.3 wt. % of the serpin in nanonized form.

The following are preferred embodiments of the NRTIs according to the invention:

An NRTI, which is

(a) suitable for local administration.

(b) selected from the group consisting of abacavir, didanosine, emtricitabine,

lamivudine, stavudine, zalcitabine and zidovudine (AZT).

(c) is nanonized with a polymer capable of forming nanop articles, preferably a linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide, analogue or derivative thereof, in particular polyhexamethylene biguanide

(PHMB). The following are preferred embodiments of the serpins according to the invention: A serpin, which is

(a) suitable for local administration.

(b) Alpha-1 Antitrypsin or ai -antitrypsin (A1AT).

(c) is nanonized with a polymer capable of forming nanoparticles, preferably a linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide, analogue or derivative thereof, in particular PHMB.

The following are preferred embodiments of the immunosuppressants according to the invention:

(a) suitable for local administration.

(b) selected from the group consisting of azathioprine, lenalidomide, methotexate, omalizumab, pomalidomide, thalidomide, prednisolone, prednisone, methylprednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide and loteprednol or a pharmaceutically acceptable salt thereof.

The following are preferred embodiments of the statins according to the invention:

(a) suitable for local administration.

(b) selected from the group consisting of atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin or a pharmaceutically acceptable salt thereof.

(c) is nanonized with a polymer capable of forming nanoparticles, preferably a linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide, analogue or derivative thereof, in particular PHMB. Particularly preferred are nanoparticles obtainable from d4T, AZT, Al AT, thalidomide prednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide, lovastatin, doxycycline or TUDCA and PHMB. Most preferred are nanoparticles formed from Al AT, AZT, thalidomide, prednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide, doxycycline or TUDCA and PHMB, optionally in the presence of a nucleonic acid, in particular adenosine triphosphate (ATP), a rhamnolipid biosurfactant and/or chitosan having a particle size determined by dynamic light scattering (DLS) in the range of about 80 nm to about 190 nm, which can be advantageously administered for the treatment of macular degeneration, stroke and COPD.

The following are preferred embodiments of the topical composition according to the invention:

A topical composition, which

(a) comprises a linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide, analogue or derivative thereof, particular PHMB, as the

nanoparticle-forming polymer.

(b) is in the form of aqueous eye drops.

(c) comprises water, a buffer and/or a stabilizer.

Most preferred are eye drops comprising nanoparticles formed from Al AT, AZT, thalidomide, prednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide, lovastatin, doxycycline or TUDCA and PHMB, water, a buffer, preferably a boric acid buffer, and a stabilizer.

EXEMPLIFICATION

The invention now being generally described, will be more readily understood by reference to the following Examples, which are included merely for purposes of illustration of certain aspects and embodiments of the present invention, and are not intended to limit the invention.

All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

Example 1 Nanonization ofNRTIs

Dynamic light scattering (DLS) is used to estimate size of complexes formed. It is based on the principle that by measuring scattered light from particles in motion their size can be determined. NRTIs (100 μg/ml) and PHMB are thoroughly mixed in w/w ratio 1 :3 in phosphate buffered saline (PBS: aqueous solution comprising NaCl, KC1, Na₂HP04, KH₂PC"4) and kept for 20 minute at room temperature. Particle size is determined using Zetasizer S (Malvern instruments, UK).

Immunosuppressants, statins, tetracyclines and bile acids in particular prednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide, thalidomide, lovastatin, doxycycline and TUDCA are nanonized analogously. Example 2 Nanonization of A 1 AT

Dynamic light scattering (DLS) is used to estimate size of complexes formed. Each formulation was prepared in water using magnetic stirring protocol, in triplicated.

Different samples (Prolastin®, Zemaira® and isolated Al AT) have been used. The formation of the nanonized form was carried out in the presence of different counter ions for the different Al AT preparation and different Al AT : Counterlon : PHMB ratios. The nanoparticle preparations displayed favorable DLS values as shown below in Table 1. TABLE 1. DLS measurements of A1AT PHMB formulations.

1 : A1AT-P: sample 1 - Prolastin®; A1AT-Z: sample 2 - Zemaira®; A1AT: sample 3 -isolated A1AT

All prepared formulations were examined to determine particle stability over time. The samples of nanonized Al AT were stored at room temperature up to 7 days and subjected to DLS measurements. The DLS measurements were performed at the following time points: 1 hour, 1 day, 3 days, 5 days and 7 days.

The results shown in Figures 1 and 2 confirm that the nanonized Al AT formulations exhibit sufficient stability. Based on the DLS values obtained (Size, the polydispersity index (PDI) and Count Rate) the best stability over time was observed for nanonized Al AT-Z (Zemaira® antitrypsin) and ATP as counter ion (Al AT-Z ATP PHMB).

In summary, nanoparticle sizes where in the range of about 80 to about 190 nm for the 3 different Al AT preparations. The PDI where acceptable for all 3 samples but were particularly good for the Zemaira® preparations and the particle counts for all 3 preparations were acceptable.

Stability of the nanoparticles was very good over 7 days for the ATP counter ion formulated AAT Z and PHMB. Example 3 Nanonization of Prednisolone

Prednisolone is nanonized as described in Example 1 of US 5,833,891 using compressed C0₂ as energizing gas and anti- solvent.

Dexamethasone, fluocinolone acetonide and triamcinolone acetonide are nanonized analogously.

Example 4 Treatment of Macular Degeneration

An aqueous ophthalmic composition is prepared comprising

0.05 % by weight of d4T, which has been nanonized as described in Example 1,

Poloxamer 407, sodium phosphate, tris-buffer and sorbitol

Said eye drops (10 drops, 4 times a day) are administered to 5 patients suffering from MD for 7 days. The treatment slows the loss of visual acuity in more than 3 patients and even increases visual acuity in 2 patients.

Example 5 In-vivo test

The cynomolgus monkey pedigree with early onset macular degeneration are tested on the effect of nanonized prednisolone and nanonized AZT

At the breeding facility, 10 female monkeys ranging from 13 to 25 year are examined. The mean age is 16.94 years, and the median age is 17 years. The clinical examination is performed by 20/30/40 drops of nanonized prednisolone as a mixture of artificial tear fluid with 2.0 g nanonized prednisolone. The cornea is kept moist with artificial tears. Fundus examination and fluorescein angiography (FA) is performed using a camera.

Each monkey is given doses of 60 mg, through two weeks, 10 mg every second day. It is to be seen that the treatment of MD with the nanonized glucocorticoid has the ameliorate effect in vision loss.

Claims

CLAIMS:

1. A topical composition for the treatment or amelioration of macular degeneration (MD), comprising one or more active ingredients, which are efficient for the treatment of MD by intraocular injection, or a pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable excipient, wherein said active ingredient is in the form of nanoparticles.

2. A topical composition according to claim 1, which comprises a polymer capable of forming nanoparticles, wherein said active ingredient is transformed into nanoparticles by said polymer.

3. A topical composition according to claim 1 or 2, wherein said one or more active ingredients are selected from the group consisting of nucleoside reverse

transcriptase inhibitors (NRTI), serine protease inhibitors (serpin), immunosuppressants, statins, tetracyclines and bile acids or pharmaceutically acceptable salts or prodrug thereof.

4. A topical composition as claimed in one of the claims 2 or 3, wherein the polymer comprises a linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide, analogue or derivative thereof.

5. A topical composition as claimed in claim 4, wherein the polymer capable of forming nanoparticles is polyhexamethylene biguanide (PHMB).

6. A topical composition as claimed in one of the claim 1 to 5, wherein the active ingredient is Alpha- 1 Antitrypsin or a 1 -antitrypsin (A1AT).

7. A topical composition as claimed in one of the claim 1 to 5, wherein the active ingredient is selected from the group consisting of abacavir, didanosine,

emtricitabine, lamivudine, stavudine, zalcitabine and zidovudine (AZT) or a

pharmaceutically acceptable salt thereof.

8. A topical composition as claimed in one of the claim 1 to 5, wherein the active ingredient is selected from the group consisting of azathioprine, lenalidomide, methotexate, omalizumab, pomalidomide, thalidomide, prednisolone, prednisone, methylprednisolone, dexamethasone, fluocinolone acetonide, triamcinolone acetonide and loteprednol or a pharmaceutically acceptable salt thereof.

9. A topical composition as claimed in one of the claim 1 to 5, wherein the active ingredient is selected from the group consisting of atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin or a pharmaceutically acceptable salt thereof.

10. A topical composition as claimed in one of the claim 1 to 5, wherein the active ingredient is selected from the group consisting of doxycycline, minocycline and tauroursodeoxycholic acid (TUDCA) or a pharmaceutically acceptable salt or prodrug thereof.

11. A topical composition as claimed in any one of the claim 1 to 10, which is in the form of aqueous eye drops.

12. A topical composition as claimed in any one of the claim 1 to 11, wherein the excipient comprises water, a buffer and a stabilizer.

13. A nucleoside reverse transcriptase inhibitor (NRTI), a serine protease inhibitor (serpin), an immunosuppressant, a statin, a tetracycline or a bile acid or a pharmaceutically acceptable salt or prodrug thereof for the treatment or amelioration of macular degeneration, wherein the NRTI, serpin, immunosuppressant, statin, tetracycline or bile acid is in the form nanoparticles.

14. An NRTI, a serpin, an immunosuppressant, a statin, a tetracycline or a bile acid according to claim 13, for local administration.

15. An NRTI according to claim 13 or 14, selected from the group consisting of abacavir, didanosine, emtricitabine, lamivudine, stavudine, zalcitabine and zidovudine (AZT) or a pharmaceutically acceptable salt thereof.

16. A serpin according to claim 13 or 14, which is Alpha-1 Antitrypsin or al- antitrypsin (Al AT).

17. An immunosuppressant according to claim 13 or 14, selected from the group consisting of azathioprine, lenalidomide, methotexate, omalizumab, pomalidomide, thalidomide, prednisolone, prednisone, methylprednisolone, triamcinolone acetonide, fluocinolone acetonide and loteprednol or a pharmaceutically acceptable salt thereof.

18. A statin according to claim 13 or 14, selected from the group consisting of atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin or a pharmaceutically acceptable salt thereof.

19. An NRTI, a serpin, an immunosuppressant, a statin, a tetracycline or a bile acid according to any of the claims 13 to 18, which is nanonized with a polymer capable of forming nanoparticles.

20. An NRTI, a serpin, an immunosuppressant, a statin, a tetracycline or a bile acid according to any of the claims 13 to 19, wherein the polymer capable of forming nanoparticles is a linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide, analogue or derivative thereof.

21. An NRTI, a serpin, an immunosuppressant, a statin, a tetracycline or a bile acid according to any of the claims 13 to 20, wherein the polymer capable of forming nanoparticles is polyhexamethylene biguanide.

22. Nanoparticles formed from Al AT, AZT, thalidomide, prednisolone, triamcinolone acetonide, fluocinolone acetonide, lovastatin, doxycycline or TUDCA and polyhexamethylene biguanide (PHMB), optionally in the presence of a nucleonic acid, in particular adenosine triphosphate (ATP), a rhamnolipid biosurfactant and/or chitosan, which exhibit a particle size determined by dynamic light scattering (DLS) in the range of about 80 nm to about 190 nm.