MXPA06004289A - Photodynamic therapy for local adipocyte reduction - Google Patents

Abstract

The present invention is drawn to methods and compounds for transcutaneous photodynamic therapy ("PDT") of target adipocyte cells or adipose tissue in a mammalian subject, which includes administering to the subject a therapeutically effective amount of a photosensitizing agent or a photosensitizing agent delivery system or a prodrug, where the photosensitizing agent or photosensitizing agent delivery system or prodrug selectively binds to the target tissue;and irradiating at least a portion of the subject with light at a wavelength absorbed by the photosensitizing agent or if prodrug, by a prodrug product thereof, where the light is provided by a light source, and where the irradiation is at low fluence rate that results in the activation of the photosensitizing agent or prodrug product. These methods of transcutaneous PDT are useful for the reduction of adipose tissue and adipocytes.

Description

PHOTODINAMIC THERAPY FOR LOCAL ADIPOSIT REDUCTION FIELD OF THE INVENTION The present invention relates in general to the field of medicine and pharmaceutics with agents of photosensitization or other agents activated by energy. Specifically, methods, compositions, compositions and equipment useful for the specific delivery at the site of a therapeutically effective amount of a photosensitizing agent for adipocytes are provided in the present disclosure. In particular, methods of use are provided, whether an external or internal light source effective to provide transcutaneous photodynamic therapy for the reduction of local adipocytes.

BACKGROUND OF THE INVENTION Obesity is a major public health problem that increases the risk of diabetes mellitus dependent on lack of insulin, stroke, heart disease, liver disease, orthopedic conditions and some types of cancer. Obesity reflects an increased volume of adiposity and an increased number of adiposities. See Prins, J. et al., Biochem. Biophys, Research Comm. 201 (2): pages 500 to 507 (1994).

Obesity is usually treated by monitoring diet, exercise and reducing subcutaneous fat layers through plastic surgery, liposuction, ultrasound and laser treatments. Due to the rapid pace of modern society, many find it difficult to maintain a healthy diet and exercise regularly in order to avoid obesity. Plastic surgery and liposuction are invasive procedures that require significant recovery periods. Invasive procedures additionally subject the patient to risks of infection, bleeding, risks from anesthesia and other post-surgical complications. Liposuction involves the introduction into the fatty layers of probes of a diameter of approximately 5 mm through holes in the skin to remove adipose tissue. The disadvantages of liposuction include the creation of a visible deficiency of homogeneity in the shape of depressions in the area of probe insertion, excessive bleeding and nonselective removal of fat and stromal cells. See Paolini et al., U.S. Patent. No. 5,954,710. The disadvantage of using subcutaneous ultrasonic samples, also includes a visible lack of homogeneity. Paolini et al., In the Pateníe of E.U.A. No. 5,954,710, describes the use of a laser beam for the removal of subcutaneous fatty layers. The described laser device comprises a needle for inserting and guiding the optic fiber that emits the laser beam into the adipose tissue to be framed. The disadvantage of using this disposition is that the radiation is invasive.

Clearly there is a need for a period of time to treat obesity by reducing adipose tissue, which method is non-invasive or minimally invasive and results in the homogeneous reduction of adipose tissue. The present invention provides a minimally invasive and non-invasive approach and method for eliminating obesity that involves the use of photodynamic therapy (PDT) to induce the reduction of adipocytes. This method and devices are described in the present description below.

BRIEF DESCRIPTION OF THE INVENTION The present invention is based on the precise direction of photosensitization agents or other agents activated by energy, drugs and compounds to the specific target cells or compositions of a subject or patient and to the acylvation period of these objective photosensitization agents or other agents. energized by mediating the administration subsequent to the subject of light or ultraphonic energy at a relatively low intensity index and over a prolonged period of time, using a light or source of ultrasonic energy that is either internal or external to the tissues objectives in order to achieve maximum cytotoxicity with minimal side effects. One embodiment includes a method for photodynamic therapy ("PDT") of subcutaneous adipose tissue in a mammalian subject comprising: administering to the subject a therapeutically effective amount of a photosensitizing agent or a system of administration of a photosensitizing agent or a prodrug, wherein the photosensitizing agent or prodrug agent or photo-sensitization agent delivery system selectively binds an objective molecule, which is an adiposity. This step is followed by the irradiation of at least a portion of the subject with light at a wavelength or wave band absorbed by means of a photosensitizing agent or if a prodrug exists, mediating a prodrug prodrug thereof, wherein the light a light source is provided, and wherein the irradiation is at a relatively low fluence index which results in the activation of the photosensitization agent or prodrug product. In this embodiment, the photosensitizing agent or prodrug agent administration or delivery system is removed from the non-target tissues of the subject prior to irradiation. Another embodiment includes a method for the transcutaneous PDT of an objective composition in a mammalian subject comprising: administering to the subject a therapeutically effective amount of a photosensitizing agent or a system of administration of a photosensitizing agent or a prodrug, wherein the agent The photosensitization or administration system of the fofosensitization or prodrug agent binds in a selective manner the objective composition. This step is followed by the irradiation of at least a portion of the subject with light at a wavelength or band of wave absorbed by the foosensitizing agent or if a prodrug exists, by a prodrug production thereof, wherein said light is provided by a light source and wherein the irradiation is at a relatively low creep index which results in the activation of the photosensitizing agent or said prodrug product. This modality contemplates that the photosensitizing agent or the administration system of the photosensitization agent or prodrug is eliminated from the non-objecid objects of the subject before said irradiation. This modality also contemplates that the light is delivered from a source of relatively non-coherent or coherent light energy that is placed close to the adipose tissue, below the surface of the skin and external to the adipose tissue. Ofra modality includes a transcutaneous PDT method of an objective tissue in a mammalian fascia as described above, wherein the light source is completely external to the intact skin layer of the patient. Another modality is directed to a method of transcutaneous PDT, where the agent of photosensitization is conjugated to a ligand. One embodiment includes a method of transcutaneous PDT, wherein the ligand is a specific antibody for adiposity and an adipocyte component, such as a lipoprotein lipase (see Sato et al., Poulíry Science 78: pages 1286 to 1291 (1999)). Other embodiments include transcutaneous PDT methods, wherein the ligand is a peptide or polymer specific for adiposifos. In certain modalities directed to a transcutaneous PDT method, the fofosensitization agent is selected from the group consisting of: green nanocyanin (ICG); Methylene blue; toluidine blue, aminolevulinic acid (ALA); phthalocyanines; porphyrins; texaphyrins; chlorine compounds; purpurins; and any other agent that absorbs light in a range of 500 nm to 1100 nm. More specifically, the chlorine and purpurin compounds contemplated in certain embodiments include: aminocarboxylic acid derivatives of cyclic or non-cyclic mono-, di- or polyamide tetraamidoes (see Bommer et al., U.S. Patent Nos. 4,675,338 and 4,693,885 , each of which is incorporated into the present description in its entirety); and pyropheoforbide-a alkyl ether derivatives with N-substituted cyclic imides (purine-18 imides) (see Pandey et al., WO 99/67249). Another modality contemplates that the photosensitization agent is mono-L-asparfil chlorine e6 (NPe6). Another embodiment includes a transcutaneous PDT method, wherein the accretion of the folosensitizing agent will likely occur within a period of from 30 minutes to 72 hours of irradiation, more preferably within 60 minutes to 48 hours of irradiation and more preferably within 3 hours. hours up to 24 hours of irradiation. Of course, clinical trials will be required to determine the optimal lighting time. Additionally, it is contemplated that the administered tofal fluence will preferably be between 30 Joules to 25,000 Joules, more preferably to 100 Joules to 20,000 Joules, and even more preferably to 500 Joules to 10,000 Joules. Clinical tests will determine the optimal total fluency required to reduce adipose tissue. A further embodiment is directed to a method for transcutaneous photodynamic therapy of objective tissue in a mammalian subject comprising: administering to the subject an effective therapeutic therapeutics of a first conjugate comprising a first member of a ligand-ligand linker pair conjugated to an antibody or antibody fragment, wherein the antibody or antibody fragment is selectively linked to an objective organism found in the adipose. This step is followed by administration to the subject of a therapeutic efficacy level of a second conjugate comprising a second member of the ligand-receptor linker pair to a photosensitizing agent or a photosensitization agent or prodrug administration system, wherein the first member is linked to the second member of the ligand-receptor linker pair. A subsequent step includes irradiating at least a portion of the subject with light at a wavelength or band of wave absorbed by the photosensitizing agent or if the prodrug exists, by the product thereof. This embodiment further includes that the light is provided by a light source and that the irradiation is at a relatively low creep index which results in the activation of the photosensitizing agent or prodrug product.

Still further embodiments refer to methods for transcutaneous PDT, wherein the ligand-receptor linker pair is selected from the group consisting of: biofin-streptavidin and antigen-antibody. A further embodiment refers to the methods described at present, wherein the antigens are adiposity antigens and the ligand-receptor linker includes biofin-sirepovidin. In this modality, the acfivation of the photosensitization agents by means of a light source of relatively low index during a prolonged period of time results in the reduction or reduction of adiposity. Another modality comprises a transcutaneous PDT method wherein the photosensitization agent administration system comprises a liposome administration system consisting essentially of the photosensitization agent. Still another embodiment includes a method for transcutaneous ulysonic therapy of an objective tissue in a mammalian fastener comprising: administering to the subject an effective epidurally effective amount of an uliosonic sensitizing agent or an ultrasonic sensitizing agent or a prodrug administration system, wherein the ultrasonic sensitizing agent or administration system of the ulimasonic sensitizing agent or prodrug is selectively linked with the adiposy. This step is followed by the irradiation of at least a portion of the fastener with ulysonic energy at a frequency that activates the ulfsonic sensitizing agent or if there is a prodrug, by a prodrug product thereof, wherein the ultrasonic energy is provided by a source of ulysonic energy emission. This modality additionally allows the ulimasonic therapy drug to be eliminated from the non-target tissues of the subject prior to irradiation. The embodiment includes a method for transcutaneous ultrasonic therapy of a target tissue, wherein the target tissue is adipose tissue. Other determined modalities contemplate that the source that emits ultrasonic energy is exíerna to the layer of iniacía skin of the patient or is inserted under the layer of iniacía skin of the patient. A further embodiment is that the ultrasonic sensitizing agent is conjugated to a ligand and more preferably, wherein the ligand is selected from the group consisting of: an adipocyte-specific antibody, an adiposite-specific peptide, and an adipocyte-specific polymer. Other modalities contemplate that the ulimasonic sensitization agency is selected from the group consisting of: green Indocyanine (ICG); methylene blue; blue of loluidine; aminolevulinic acid (ALA); phthalocyanines, porphyrins; texaphyrins; pyropheophorbide compounds; Chlorine compounds; purpurins; and any other agent that absorbs light in a range of 500 nm to 1100 nm. More specifically, the chlorine and purpurin compounds contemplated include: aminodicarboxylic acid derivatives of cyclic or noncyclic mono-, di-, or polyamide (see Bommer et al., U.S. Patent Nos. 4,675,338 and 4,693,885); and pyropheoforbide-a alkyl ether derivatives with N-substituted cyclic me mides (purine-18 midas) (see Pandey et al., WO 99/67249). One embodiment contemplates that the photosensitizing agent is mono-L-aspartyl chlorin e6 (NPe6). Other embodiments include the currently described methods of transcutaneous PDT, wherein the light source is positioned in proximity to the target tissue of the subject and is selected from the group consisting of: a LED light source; a source of electroluminescent light; an incandescent light source; a light source of organic polymer and an inorganic light source. One mode includes the use of an LED light source. Still other embodiments of the currently described methods are directed to the use of light of a wavelength that is from about 500 nm to about 1100 nm, preferably greater than about 650 nm and more preferably greater than about 700 nm. One embodiment of the present method is directed to the use of light which results in a single photon absorption mode by the photosensitizing agent. Additional embodiments include targeted photosensitization delivery system compositions comprising: a photosensitizing agent and a ligand that binds to a receptor in the target tissue with specificity. In one modality, the photosensitization agent of the directed administration system is conjugated to the ligand that links a receptor on the objective lesion with specificity. Preferably, the ligand comprises an antibody that binds to a receptor and the receptor is an antigen on adipocytes. Still more preferred is the lipoprotein lipase anígeno, which binds specifically and preferentially to the monoclonal antibodies of lipoprotein lípase (see Sato ei al., Poulfry Science 78: pages 1286 to 1291 (1999)). An additional embodiment contemplates that the photosensitizing agent is selected from the group consisting of: green indocyanine (ICG); methylene blue: toluidine blue; aminolevulinic acid (ALA); phthalocyanines; porphyrins, texaphyrins, chlorine compounds; purpurins; and any other agent that absorbs light within a range of 500 nm to 1 100 nm. Another embodiment of the present invention contemplates that the photosensitizing agent is mono-L-asparyl chlorin e6 (NPe6). Still another modality includes the ligand-receptor binding pair that is selected from the group consisting of: biotin-streptavidin and antigen-host. Still another embodiment contemplates that the photosensitizing agent comprises a prodrug. Other embodiments contemplate methods for transcutaneous PDT to deplete an objective cell in a mammalian subject comprising: administering to the subject a therapeutically effective amount of a photosensitizing agent or a system of administration of photosensitizing agent or a prodrug, wherein the agent The photosensitization or administration system of the foesensitization or pro-drug agent is selectively linked to the target cell. This step is followed by the irradiation of at least a portion of the holder with light at a wavelength or band of wave absorbed by the photosensitizing agent or if the prodrug exists, by a prodrug production thereof, wherein the light it is provided by a light source, and wherein the irradiation is at a relativly low creep index which results in the activation of the photosensitizing agent or prodrug product and the desolution of the target cell. This modality contemplates that the photosensitization agent is cleared of the non-objelive objects of the subject before said irradiation. Still a further embodiment allows a photosensitizing agent to be delivered locally or regionally by administering a drug delivery patch method. This modality is also provided for the use of ultrasound to boost and direct the agent of foiosensibilization in the subcutaneous fatty acids. An alternative methodology is provided for the percutaneous injection of the trainer's site.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows a diagram demonstrating transcutaneous PDT that uses a laser diode light source that is focused (3) and not focused and placed at an angle (2) of adipose tissue (5) that is exoerne to the layer of leather (4).

Figure 2 shows the PDT using an optical fiber (6) that administers light from a laser diode light source (2) that is inserted under the skin layer (4), although it is external to the outer membrane of the skin. adipocyte (5). Figure 3 shows the transcutaneous PDT that uses a light source that is comprised of multiple LEDs arranged on a band (7) or a fiber optic diffuser (7) and is placed outside the skin layer (4). Figure 4 shows the transcutaneous PDT using an optical diffuser (8) attached to an optical fiber with administration of light from a laser diode light source (not shown). Figure 4A shows an end of the view of the optic fiber with a reflected surface (9) that directs the light towards the radiating area.

DETAILED DESCRIPTION OF THE INVENTION Apoptosis is a specific form of cell death. Apoptosis occurs under normal conditions, such as during embryogenesis and physiological involution of adult tissue. This also occurs during abnormal conditions or can be induced by exposure to radiation, neoplastic drugs and other toxins. It has been suggested that apoptosis may play a role in the reduction of adipocytes. See Prins, J. et al., Diabetes 46: pages 1939 to 1944 (1997)), and Prins, J. et al., Biochem. Biophys. Research Comm. 205 (1): pages 625 to 630 (1994).

One form of energy-derived therapy is the foodyodynamic therapy (PDT). PDT has been applied to treat a range of conditions including cancer and heart disease. See Oleinick, N. et al., Radiation Research 150: pages S146 to S156 (1998). PDT can be used to induce apoptosis. See Ahmad, N. et al., Proc. Nati Acad. Scí. 95: pages 6977 to 6982 (1998); and Kessel, D. et al., Cell Death and Differentiation 6: pages 28 to 35 (1999). The PDT is performed by first administering a pseudosensitization compound in a systèmic or isopic form, followed by illumination of the irradiation site at a wavelength or wave band, which closely matches the spectroscopy of the photosensitizer. By doing so, the oxygen singlet and other reactive species are generated leading to a number of biological effects that result in cytotoxicity. The depth and volume of the cytotoxic effect on the tissue depend on the complex interactions of the light penetration in the tissue, the concentration of the photosensitizer and the cellular location, and the availability of molecular oxygen. A large number of PDT light sources and methods of use have been described. However, the reports describing the sources and effects of transcutaneous light administration for the purposes of the PDT are more limited. It has generally been accepted that the ability of a light source external to the body to produce clinically useful loxiciency is limited in depth to a depth of 1 to 2 cm, or less depending on the photosensitizer. Accordingly, the gradual reduction of subcutaneous adipose tissue can occur in a non-invasive manner without causing extensive tissue damage. The methods, compounds, compositions and equipment described in the present description maintain that PDT will be used to induce apoptosis instead of necrosis of adipocytes. By administering a therapeutically effective concentration of the photosensitizer or acyivative agent by energy and modulating the amount of irradiation energy, the degree of necrosis and subsequent inflammation can be minimized. Additionally, this will ensure that other side effects due to rapid triglyceride mobilization can be avoided or decreased. The apoptotic process allows a much more controlled reduction of fatty tissue deposits compared to a procedure in which, said tissue is reduced by an induction of cell necrosis. However, the treatment of subcutaneous fat layers, in this way, can be associated with inadvertent skin damage due to the accumulation of the photosensitizer in the skin, which is a property of all sensitizers administered systemically in the skin. clinical use For example, clinically useful porphyrins, such as Photorphrin® (QLT, Ltd. Sodium porfimer label) are associated with photosensitivity that lasts up to 6 weeks. Purlyfin®, which is a glitter and Fosean®, which is a chlorine, sensitizes the skin for several weeks. In fact, efforts have been made to develop photoresists to reduce skin photosensitization (see Dillon et al., Photochemistry and photobiology 48 (2): pages 235 to 238 (1988)).; and Sigdestad et al., British J. of Cancer 74: pages S89 to S92 (1996)). In fact, PDT protocols involving the systemic administration of the photosensitizer require the patient to avoid sunlight and bright indoor light to reduce the possibility of phototoxic reactions of the skin. One modality of the PDT describes the use of an intense laser light source to activate the drug within a precisely defined limit. See Fischer et al., Patent of E.U.A. No. 5,829,448. A two-photon methodology requires high-power laser light for drug activation with a highly collimated beam that requires a high degree of spatial control. This type of irradiation is not practical for fraying large areas of adipose tissue, because the ray may need to be passed quickly through the surface of the skin in some type of group, with a pattern that can be repeated by time. Movement of the patient or organ could be a problem due to misalignment. The tissue or skin not targeted and the photosensitivity of the subcutaneous tissue is not addressed in the available literature. Any fofosensitizer in the irradiation of the ray could be activated and produce unwanted collateral tissue damage. Therefore, a one-photon method is preferred in PDT reduction of adipose tissue. The one-photon method allows prolonged exposure to a lower creep index, which promotes the protection of non-target tissue or skin and normal subcutaneous tissue, and reduces collateral tissue damage. The present invention further describes the selective binding of the photosensitizing agent to specific target tissue antigens, such as those found on the surface of or within the adipocytes. This targeting scheme decreases the amount of sensitization drug required for effective therapy, which in turn decreases the total fluence, and the fluence index necessary for effective photo-activation. A much less intense light source is preferred than a high power, short exposure laser beam that uses collimated light as described by W.G. Fischer et al., In Photochemistry and photobiology 66 (2): pages 141 to 155 (1997). The present invention allows the use of a non-coherent light source of low power used for a period longer than one hour to increase the depth of foloactivation. The present invention provides methods and compositions for the treatment of an objective tissue or the removal or deterioration of the target cell or composition in a mammalian subject by specific and selective binding to the target tissue, cell or composition of a photosensitizing agent. This method comprises irradiating at least a portion of the subject with light at a wavelength absorbed by said photosensitizing agent which under activation conditions lasts photodynamic therapy using a relatively low creep index, although a high total fluence dose in General results in minimal collateral tissue damage. The terms as used in the present description are based on their recognized meaning in the subject and of the present description that must be clearly understood by a person ordinarily skilled in the maize. For the purpose of clarity, the terms may also have a particular meaning that could be clear from their use in the context. For example, the term transcutaneous refers in the present description more specifically to the passage of light through non-open tissue. Where the tissue layer is the skin or dermis, transcutaneous is transdermal and the light source is extruded to the outer skin layer. However, in cases of transillumination, the description refers to the passage of light through a layer of light.As a layer of adipose tissue, the source of light is excreted to the adipose tissue, although internal or implanted in the subject or patient. Specifically, the present embodiments are based on the precise location of photosensitizing agents or drugs and compounds for specific target antigens of a subject or patient and the method of activating localized photosensitization agents by subsequent administration to the light subject in a Relatively low creep index during a prolonged period of time from a light source that is external to the objective material with the objective of achieving maximum cyto-toxicity or reduction of adipocytes over time with minimal side effects or collateral tissue damage. Additionally, as used herein, "target cells" or "target tissues" are those cells or tissues, respectively, that are intended to be deodorized or destroyed by this method of treatment. Target cells or target tissues use the photosensitizing agent; then when sufficient radiation is applied, these cells or tissues are deteriorated or desiruded. The objective cells are those cells in the target tissues, which include, but are not limited to, adiposy or preadiposy. The "non-objective cells" are all the cells of an intact animal that are not intended to be damaged or destroyed by the treatment method. These non-target cells include, but are not limited to, stromal cells, and other normal tissue, which are otherwise not identified to be localized. The term "destroy" is used to understand to annihilate the target cell. The term "deteriorate" means to change the target cell in such a way that it interferes with its function. For example, North et al., Observed that after exposure to the light of T cells infected with virus, frayed with benzoporphyrin derivatives ("BPD"), holes were developed in the T cell membrane, which were increased in size until the membrane completely decomposed (Blood cells 18: pages 129 to 140 (1992)). The target cells are understood to be impaired or desirmed even if the target cells are arranged in the last insanity by the macrophages. The term "photosensitization agent" is a chemical compound, which is housed in one or more selected target cell types and when it is placed in coniaction by radiation, it absorbs light, which results in the deterioration or destruction of the cells objective. Virtually any chemical compound that is housed in a selected target and absorbs light can be used by the present invention. Preferably, the chemical compound is non-toxic to the animal in which it is administered or has the ability to be formulated in a non-toxic composition. Preferably, the chemical compound in its fode-degraded form is also non-toxic. A broad list of photosensitive chemicals can be found in the Kreimer-Brinbaum publication, Sem. Hemalol. 26: pages 157 to 173 (1989). Photosensitive compounds include, but are not limited to: green Indocyanine (ICG); methylene blue; oluidine blue, aminolevulinic acid (ALA); phthalacyanines, porphyrins; texaphyrins; bacteriochlorins, merocyanines, psoralens, benzoporphyrin derivatives (BPD) and sodium porfimers and prodrugs such as d-aminolevulinic acid, which can produce drugs such as protoporphyrin. Also included are: chlorine compounds, purpurins and any other agent that absorbs light within a range of 500 nm to 1100 nm. More specifically, the chlorine and purpurin compounds contemplated by the present invention include: aminodicarboxylic acid derivatives of cyclic or noncyclic mono-, di-, or polyamide tetrapyrroles (see Bommer et al., US Patent Nos. 4,675,338 and 4,693,885); pyropheoforbide-a alkyl ether derivatives with N-substituted cyclic imides (purine-18 imides) (see Pandey et al., WO 99/67249). Specifically, derivatives of mono-L-aspartyl chlorin e6 (NPe6) and any other agent that absorbs light within a range of 500 nm to 1100 nm are included. The term "radiation" as used in the present description includes all wavelengths. Preferably, the wavelength of radiation is selected to coincide with the wavelength (s) that mimic the photosensitive composite. Still more preferably, the wavelength of radiation coincides with the stimulation wavelength of the photosensitive compound and has low absorption of the non-target cells and leave the rest of the animal intact, including the blood proteins. For example, the preferred wavelength for conventional NPe6 is within the convenient range of 600 to 800 nanometers, with preferred compounds being absorbed within a range of 620 to 760 nanometers. Radiation is defined additionally by its intensity, duration and registre of time with respect to the dosage with the photosensitive agent. The intensity or creep index must be sufficient for the radiation to penetrate the skin and reach the target cells, target tissues or target compositions. The duration or total fluence dose should be sufficient to photo-activate the photosensitive agent sufficient to adhere to the target cells. Both intensity and duration should be limited in order to avoid animal over-exposure. The time record with respect to dosing with the photosensitive agent is important because (1) the agent of administered photosensitization requires some time to lodge in the cells and (2) the blood level of many photosensitization agencies decreases rapidly over time. The present invention provides a method for treating an animal, which includes, but is not limited to, humans and other mammals. The term "mammals" or "mammalian subject" also includes farm animals, such as cows, pigs and sheep, as well as pets or animals for sports, such as horses, dogs and cats. By "intact animal" is intended to mean the entire animal, without dividing it, which is available to be exposed to radiation. No part of the animal is removed for separate radiation. It is not necessary that the animal as a whole be exposed to radiation. Only a portion of the iniacious animal subject can or requires exposure to radiation. The term "transcutaneously" is used in the present description as a meaning of the skin of an animal subject. Briefly, the animal sensitization agent is usually administered to the animal before the animal is subjected to radiation. Preferred photosensitizing agents include, but are not limited to: green indocyanine (ICG) (e.g., see WO 92/00106 (Raven et al); WO 97/31582 (Abis et al.) And Devoisselle et al., SPIE 2627: pages 100 to 108 (1995)); Methylene blue; toluidine blue; and prodrugs, such as delta-aminolevulinic acid, which can produce drugs such as protoporphyrin; bacteriochlorins; phthalocyanines; porphyrins; texaphyrins; chlorine compounds; purpurins; merocyanins; psoralens, and any other agent that absorbs light within a range of 500 nm to 1100 nm. More specifically, the chlorine and purpurin compounds contemplated in the present invention include aminodicarboxylic acid derivatives of cyclic and non-cyclic mono-, di-, or polyamide tetramers (see Bommer et al., U.S. Patent Nos. 4,675,338 and 4,693,885. ); and pyropheoforbide-a alkyl ether derivatives with N-substituted cyclic imides (purpurin-18 midas) (see Pandey et al., WO 99/67249). An additional foesensitizing agent is mono-L-aspartyl chlorin e6 (NPe6) (see U.S. Pat. No. 4,693,885). The photosensitizing agent is administered locally or systemically. The photosensitizing agent is administered orally or by injection, which may be iniravenous, subcutaneous, intramuscular or intraperitoneal. The photosensitizing agent can also be administered externally or topically by means of patches or implants. The photosensitizing agent can also be conjugated to specific ligand reagents with a target, such as receptor-specific ligands or immunoglobulins or immunospecific portions of immunoglobulins, allowing them to run more concentrated in a desired target cell or microorganism. The photosensitivity library can additionally be conjugated to a ligand-receptor binding pair, which includes, but is not limited to, biotin-streptavidin and antigen-antibody. This conjugation can make it possible to decrease the level of dose required because the material is directed more selectively and is less wasted in the distribution within other tissues whose destruction can be avoided. The photosensitization agent, in one embodiment, can be formulated in suitable pharmaceutical preparations, such as solutions, suspensions, iables, dispersible tablets, pills, capsules, powders, formulations or sustained-release elixirs, for oral administration or in solutions or sterile suspensions for parenferal administration, as well as in fransdermic patch preparations and dry powder inhalers. In one embodiment, the compounds described above are formulated in pharmaceutical compositions using techniques and procedures well known in the art (see, for example, Ansel, Introduction to Pharmaceutical Formulations, fourth edition, page 126, 1985). The foesensitization agent can be administered in a dry formulation, such as iables, pills, capsules, powders, granules, suppositories or patches. The photosensitizing agent can also be administered in a liquid formulation, either alone with water or with pharmaceutically acceptable excipients, such as those described in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, 15th ed. 1975. The liquid formulation can also be a suspension or an emulsion. In particular, liposomal or lipophilic formulations are more desirable. If suspensions or emulsions are used, suitable excipients include water, saline, dextrose, glycerol and the like. These compositions may contain minor amounts of non-toxic auxiliary substrates such as wetting agents or emulsifiers, antioxidants, pH regulating agents and the like. The dose of photosensitizing agent will vary with the intended target cell (s), the optimum blood level (see Example 1), the weight of the animal and the time recording of the radiation. Depending on the photosensitizing agent used, an equivalent optimal epigraphic level will have to be established. Preferably, the dose is calculated to obtain a blood level between about 0.001 and 100 μl. Preferably, the dose will obtain a blood level of about 0.01 and about 10 μl. This method comprises irradiating at least a portion of the holder with light at a wavelength or band of wave absorbed by said photosensitizing agent which, under activation conditions during photodynamic therapy, uses a relatively low creep index, but also at a dose of overall high total fluence that results in minimal collateral tissue damage. It is contemplated that the optimum total fluence will be determined clinically using a light dose increase assay. Additionally it is contemplated that the total creep will preferably be within the range of 30 to 25,000 Joules / cm 2, and more preferably will be within the range of 100 to 20,000 Joules / cm 2, and more preferably will be in the range of 500 to 10,000 Joules / cm 2. The methods comprise irradiating at least a portion of the subject with light at a wavelength or wave band absorbed by said photosensitizing agent which under conditions of activation during photodynamic therapy use a relatively low creep index, although a flow rate to high overall, resulting in minimal collateral normal tissue damage. What is meant by "relatively low creep index" is a creep index that is lower than that normally used and one that generally does not result in damage to the collateral tissues or non-objecivo. Specifically, the intensity of radiation used to irradiate the objective cell or objective tissue is preferably about 5 to 100 nW / cm 2. More preferably, the radiation intensity is between about 10 and 75 mW / cm2. More preferably, the radiation intensity is between about 15 and 50 mW / cm2. The duration of exposure to radiation is preferably within a range of between 30 minutes and 72 hours. More preferably, the duration of exposure to radiation is about 60 minutes and 48 hours. More preferably, the duration of exposure to radiation is between about 2 hours and 24 hours. Of course, routine clinical tests will be useful in determining the optimal flow rate and total fluence delivered to the treatment site. While not wishing to be bound by a theory, it is proposed in the present description that a photosensitizing agent can be substantially and selectively photoactivated in target cells and target tissues within a reasonable therapeutic time period and without excess toxicity or collateral damage. to non-objelive tissues. Therefore, it seems to be a therapeutic window limited by the dose of photosensitive agent and the radiation dose. The formation of photodegradation products of a photosensitizing agent was used as an indicator of photoacíivación. The foioacíivación of a photosensitizing agent has been postulated to produce the formation of oxygen singlet, which have a cyioxic or apoptotic effect. Additionally, certain embodiments are directed to methods for transcutaneous ulcerative therapy of adipose tissue in a mammalian subject or patient, by first administering to the subject a therapeutically effective amount of a first conjugate comprising a first member of a ligand-receptor binding pair conjugated to an antibody or antibody fragment, wherein said antibody fragment or antibody is selectively linked to a target adipocyte antigen; and in a simulative or subsequent manner, administering to the subject a therapeutically effective amount of a second conjugate comprising a second member of the ligand-receptor binding pair conjugated to an ultrasonic sensitizing agent or delivery system or ultrasonic sensitizing agent or prodrug, where the first member binds to the second member of the ligand-receptor binding pair. These steps are followed by the irradiation of at least a portion of the subject with energy at a wavelength absorbed by said ultrasonic sensitizing agent or the ultrasonic sensitization agent delivery system, by the product thereof, wherein said energy is provided by an energy source that is external to the clamp; and wherein said ultrasound is at a relatively low intensity index which results in the acyivation of said ulysonic sensitization agent or prodrug product. Although one embodiment is directed to the use of light energy in a photodynamic adipose tissue therapy utilizing light and photosensitizing agents, other forms of energy are within the scope of the present invention and are understandable by a person ordinarily skilled in the art. Said forms of energy include, but are not limited to: thermal, sonic, ulysonic, chemical; photo or light; microwave; ionization, such as: X-rays and gamma rays; and electric. For example, agents induced or activated in sonodynamic form include, but are not limited to: gallium-porphyrin complex; and other porphyrin complexes, such as protoporphyrin and hematoporphyrin. See the publication of Yumita et al., Cancer Letters, 112: pages 79 to 86, 1997; and Umemura et al., Ultrasonics Sonochemislry 3: pages S187 to S191 (1996). This modality additionally contemplates the use of an energy source that is external to the target tissue. Target tissues may include and may be related to adipocytes, per se. Those skilled in the art will be familiar with various ligand-receptor binding pairs that include those known and those that are yet to be discovered. Those known, include, but are not limited to, the group consisting of: biotin-streptavidin and ani-antigen. The present invention contemplates a modality that includes the use of biotin-streptavidin as the ligand receptor pair. However, one skilled in the art will readily understand from the present disclosure that any ligand-receptor binding pair can be useful as long as the ligand-receptor binding pair demonstrates a specificity for binding by the ligand to the receptor and additionally provided that the ligand-receptor binding pair allows the creation of a first conjugate comprising a first member of the ligand-receptor binding pair conjugated to an antibody or antibody fragment, wherein said antibody or antibody fragment binds selectively a target antigen of the adipocytes; and additionally allows the creation of a second conjugate comprising a second member of the ligand-receptor binding pair conjugated to an energy sensitizing agent or foosensitization or administration system of energy sensitizing agent or photosensitization or prodrug, and adidonally in where the first member binds to the second member of the ligand-receptor binding pair. Another group of ligand-receptor pairs includes the conjugation of an energy sensitizing or photosensitizing agent or system of administering photosensitizing agent or prodrug to a first member of the ligand-receptor binding pair selected from the group consisting of an antibody to a adipocyte-specific antigen; a ligand that can bind to an adipocyte cell receptor; and other ligands that can bind to cell surface components of specific adipocytes. Said first pair of ligand-receptor member will bind selectively and specifically to the second member of the receptor ligand binding pair, which may be an adiposite-specific adipocyte-specific antigen or other adipocyte-specific cell surface component. In this way, an agent that is acylated by energy is administered specifically to its adipocyte target cell corresponding to the selected ligand-receptor binding pair. For example, the monoclonal antibody directed against the lipoprotein lipase antigen binds specifically and preferably to the lipoprotein lipase (see, Sato et al., Poultry Science 78: pages 1286 to 1291 (1999)). Ofra modality is directed to a method wherein the administration system of photosensitization agent includes a liposome delivery system which consists essentially of the folosensitizing agent, however, the experts in the material will easily understand from the present description that other administration systems can be used. In one embodiment, liposome suspensions, which include liposomes directed to tumors, such as tumor-directed liposomes, may also be suitable as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art. For example, liposome formulations can be prepared as described in the USPatent. No. 4,522,811. Yet a further embodiment contemplates the described method wherein the administration system of the photosensitization agent uses both a liposome delivery system and a photosensitization agent, wherein each is conjugated separately to a second member of the ligand binding pair. -receptor, and wherein the first member is linked to the second member of the ligand-receptor binding pair, and more preferably wherein the binding pair of receptor ligand is biotin-sirepfavidin. This embodiment additionally contemplates that the photosensitizing agent, as well as the photosensitizing agent delivery system, can both be targeted specifically through the selective binding system to a specific tissue antigen by the antibody or antibody fragment of the first member of the link pair. This double direction is envisioned to improve the extraction specificity and to increase the exfusion rate.

Having generally described the present invention, it will be more readily understood by reference to the following examples, which are provided as an illustration, and are not intended to limit the present invention, unless so specified.

EXAMPLE 1 Transcutaneous photodynamic therapy of adipose tissue A photosensitizer can be administered in a sysiemic or regional manner. In the case of seismic administration, the foesensitizer is conjugated with an agent that allows the selective extraction of adipose tissue or adipocytes. In the case of regional administration, the photosensitizer can be administered topically. Iopy administration can be followed by a method, such as ultrasound, which improves skin impregnation and localization within the subcutaneous fat. Alternatively, the photosensitizer can be injected percutaneously into the trailing site where diffusion occurs and allows the dispersion of the photosensitizer. The process of photoactivation that is preferred is that which induces apoptosis and not the necrosis of the adipocytes. Esio reduces inflammation and other secondary effects due to the rapid mobilization of triglycerides. The apoptotic process allows a controlled reduction of adipose tissue compared to a process, by which, necrosis occurs. Triglycerides within the adipocytes subjected to PDT are released and metabolized gradually by some surrounding cells. Apoptosis can be determined in tissue explants by observing the "staggering" characteristic after gel electrophoresis, which confirms the occurrence of specific endonuclease-induced DNA separation, chromatin agglutination and lipid-laden interstitial macrophages. A. Adipocytes and adipose tissue can be effectively diminished by transcutaneous photodynamic therapy. A localized antibody-photosensitizer conjugate (APC) is prepared by linking a photosensitizing agent, such as NPe6, to a monoclonal antibody that binds to an adipocyte-specific antigen, such as a lipoprotein lipase. This APC is administered to the language center by any number of means available to an expert in the field. For example, APC can be administered by local injection under the subcutaneous skin layer or systemically by intravenous injection. The administration of other APC formulations may include: oral or topical formulations. Elstrom et al., In the U.S. Patent. No. 5,999,847 teach the use of localized and transient pressure waves that are applied to the adjacent adjoining target cells by means of a light source and a coupling interface placed in contact with the liquid that converts light from the light source into acoustic energy The pressure waves produce the formation of transient pores of cell membranes. The therapeutic agents are administered to the site of the pressure waves located by any suitable means, such as by injection with a needle. The light source and coupling interface can be incorporated into a catheter for the application of pressure waves to diseased blood vessels. A manually operable surgical device incorporating a needle for injecting the agent, light source and coupling interface can also be used. The excess of photosensitizing conjugates is eliminated in a nalural way by the body. One or more sources of light are placed or implanted in a spiral form near the tissue to be irradiated. After a sufficient amount of time to allow the clearance of non-target tissue conjugates, at about 6 hours, the light sources are positive, irradiating the target tissue at a relativly low creep rate of about 50 mW / cm2 lasts 5 hours, which results in a high total fluence dose of light, such as 900 Joules / cm2, at the wavelength from about 620 nm to about 760 nm. Light can be applied internally or externally. The specific dose of photosensitizer conjugate is that which results in an active NPe6 concentration sufficient to obtain a blood level between about 0.001 and 100 μg / ml. and more preferably, a dose of between about 0.01 and 10 μg / ml. However, it is within the abilities of a person skilled in the art to determine the specific therapeutically effective dose using standard clinical practices and procedures. Similarly, the specific creep index and the total creep dose can be determined routinely from the present disclosure. Additionally, the above conjugate could be further conjugated to an imaging agent, such as a technetium. Accordingly, the method could further comprise the steps of conducting a nuclear medical scan and imaging the sites to be brought. B. Alternatively, after the description of Example 1A, an APC can be constructed by linking a photosensitizing agent that selectively binds to a second antigen, different from the lipoprotein lipase and which is mainly present or is associated with adipocytes. The photosensitizer could, instead, be linked to a receptor-ligand link pair, where one of the link pair is specifically associated with adipocytes and the other of the link pair is linked to the photosensitizing agent. Such receptor-ligand binding pairs could include: hormone-hormone receptor, chemokine-chemokine receptor; or another signal transduction receptor and its natural ligand. The ligand-receptor binding pair or APC is implanted intravenously and is concentrated in the adipose tissue. When it is unlinked, the APC is removed from the body. Internal or external light sources can be used to activate the targeted drug, however, in this Example, an extern light source is contemplated. Any number of antigen components or ligand binding pair can be selected, provided that the component is specifically associated with the adipocytes. Such antigen components or ligand binding pair could be known to those skilled in the art. The selection of a specific photosensitizing agent can be performed, provided that the photosensitizing agent is activated by a wavelength from 500 nm to 1100 nm, and more preferably a wavelength of 620 nm, and still more preferably by a length of wave of 700 nm or greater. Said photosensitizing agents that are provided in the present description are contemplated to be used in the same. C. After the description of Example 1A and 1B above, the PDT light source is a light source placed externally connected (1) to a power source and directed to the sifio to be trafficated. The light source can be a laser diode, a light emitting diode (3) or another electrofroluminescent device. The light source can be at an angle (2) or placed perpendicular (3) to the skin layer (4) and the light beam is focused in such a way that light is directed through the skin or membrane of the skin. I hold a mammal that is being traced to produce the fof oaacíi emptiness of the photosensitizing agent linked to the adipocytes (5) of the adipose tissue. See Figure 1.

Alternatively, the light source could comprise a band or panel of light emitting diodes (LEDs) (7), which are then disposed on the skin or upper membrane of the site to be irradiated in the mammalian subject. See Figure 3. The light source could also comprise a fiber optic diffuser (8), which is placed on the skin or upper membrane of the synovium to be irradiated in the mammalian subject. Said diffuser may additionally comprise a reflected surface (9) which directs the light beam to the target area. See Figure 4. D. As is clear to one skilled in the art, the methods and compositions described above have various applications. For example, a small area of adipose tissue in a mammalian subject may be brought using a patch composed of LEDs or a woven optic fiber mat where the patch or light source mat is placed on the skin or upper tissue of the site. be irrational. Additionally, the patch or pad could also contain pharmaceutical compositions or the photosensitizing agent, which is then administered by means of liposome, fransdermic or ytophoretic techniques.

EXAMPLE 2 Photodynamic translucent therapy of adipose tissue The following is the method of Example 1A, wherein a conjugate is formed between NPe6 and the monoclonal antibody to lipoprotein lipase. Said conjugate is administered in the manner described in Example 1A. An internal light source is provided in a surgical form by an invasive procedure in minimal form. The LED (2) is connected to an optical fiber (6) and surgically inserted under the subcutaneous layer of tissue (4). For example, Chen et al., In the Pateníe of E.U.A. No. 5,766,234, teach the implantation of a fiber optic fiber with an LED light source for photodynamic therapy at a local site. Also, Paolini et al., In the U.S. Patent. No. 5,954,710 teach a device and method for removing the sub-fatty adipose layers by using a laser light source connected to an optical fiber that transports the media and a hollow needle to guide the fiber, wherein said fiber closes in the vicinity of the fiber exfiltration. needle. The present invention has been described by direct description and by examples. As noted above, the examples are intended only to be examples and not to limit the present invention in any meaningful sense. Additionally, an expert in the matter to which the present invention belongs, when reviewing the specification and claims that will be found below, will appreciate that there are equivalents for those claimed aspects of the present invention. The inventors intend to cover said equivalents within the reasonable scope of the claimed invention. The citations to the above documents are not intended as an admission that any of the above is an appropriate prior art. All statements to date or representation as content of these documents are based on information available to applicants and do not constitute an admission of correction of the dates or contents of these documents. Additionally, all documents referred to through this application are incorporated in their identity as a reference to the present description.

Claims (23)

NOVELTY OF THE INVENTION CLAIMS

1. - The use of a photosensitizing agent or a system for administering a photosensitizing agent or a prodrug, wherein said photosensitizing agent or said administration system of a photosensitizing agent or said prodrug, is selectively located in the adipose tissue or the adips; for preparing a medicament for the reduction of adipose tissue or adipocytes in a mammalian subject, and wherein the light is administrable at a relatively low fluence index which results in the activation of said photosensitizing agent or said prodrug production and wherein said PDT drug is removed from the skin and subcutaneous tissues of the subject before said irradiation, before said light is administrable.

2. The use claimed in Claim 1, wherein said source of light is selected from the group consisting of one or a plurality of: laser diodes; light-emitting diodes; sources of electroluminescent light; incandescent light sources; cold cathode fluorescent light sources; light sources of organic polymer; or inorganic light sources.

3. The use claimed in claim 1 or claim 2, wherein said source of light is exíerna to the skin layer and the ray of light is directed through the skin to adipose tissue or adiposity.

4. - The use claimed in Claim 2, wherein said laser diode is coupled to an optical fiber, and wherein said optical fiber directs said light to adipose tissue or adipocytes.

5. The use claimed in Claim 2, wherein said light emitting diode is a light emitting diode band, and wherein said light emitting diode band is located external to the skin layer and above. of adipose or adipose tissue.

6. The use claimed in claim 4, wherein said optical fiber diffuses said light when it is placed on adipose tissue or adipocytes.

7. The use claimed in Claim 4 or Claim 6, wherein said light source is a mat comprising a plurality of said optical fiber.

8. The use claimed in any of the preceding Claims, wherein said photosensitizing agent is selected from the group consisting of: green indocyanine; mephylene blue; oluidine blue, delia-aminolevulinic acid; protoporphyrin; bacoferiochlorins; flalocyanines; porphyrins; texaphyrins; merocyanines; psoralens; pyropheoforbides; chlorines; purpurins; and any other particle that absorbs light in an inervance of 500 nm at 1100 nm.

9. The use claimed in Claim 8, wherein said fofosensibilization agent is a derivative of aminocarboxylic acid of mono-, di- or polyamide of a cyclic or non-cyclic tetrapyrrole.

10. - The use claimed in any of Claims 1 to 9, wherein said photosensitizing agent is mono-L-aspartylchlorin e6 (NPe6).

11. The use claimed in Claim 1, wherein said wavelength is from about 500 nm to about 1100 nm.

12. The use claimed in Claim 1 or Claim 11, wherein said wavelength is greater than about 700 nm.

13. The use claimed in any of the Claims 1 to 12, wherein said light results in a single photon absorption mode by the photosensitizing agent.

14. The use claimed in claim 8, wherein a complex, comprising said photosensitizing agent, is conjugated with a specific ligand of adipose tissue which is located in adipose tissue or adipocytes.

15. The use claimed in Claim 14, wherein said ligand is an: adipocyte antigen; adipocyte cell receptor; or another adipocyte cell surface component.

16. The use claimed in Claim 15, wherein said antigen is lipoprotein lipase.

17. The use claimed in Claim 14, wherein said complex is administrable in a systemic or local manner.

18. The use claimed in Claim 17, wherein said complex is formulated for administration orally, topically, intravenously or by any percutaneous injection route.

19. The use claimed in Claim 17, wherein the complex is allowed to permeate the skin and the interior of the subcutaneous adipose tissue.

20. The use claimed in any of Claims 1 to 19, wherein the reduction of adipose tissue or adipocytes occurs through the apoptosis of adipocytes.

21. An apparatus for transcutaneous photodynamic therapy of adipose tissue or adiposity in a mammalian subject that comprises a light source that is external to the subject and is selected from the group consisting of one or a plurality of: laser diodes; light-emitting diodes; Electroluminescent light sources; incandescent light sources; cold cathode fluorescent light sources; light sources of organic polymer; or inorganic light sources.

22. The apparatus according to claim 21, further characterized in that said light source is at least one laser diode coupled to an optical fiber, which directs said light to adipose tissue or adipocytes.

23. The apparatus according to claim 21 or claim 22, further characterized in that said diode is a light emitting diode band, and wherein said light emitting diode band can be placed on the skin to outline the adipose tissue to be frayed.