MXPA02011638A - Halogenated rhodamine derivatives and applications thereof. - Google Patents

Abstract

Novel compounds of the formula (I) wherein: one of R1, R2, R3, R4, and (R10)n represents an halogen atom and each of the remaining R1, R2, R3, R4, and each of the remaining R10 group is independently selected in the group constituted by hydrogen, halogen atoms, an amino, acylamino, dialkylamino, cycloalkylamino, azacycloalkyl, alkylcycloalkylamino, aroylamino, diarylamino, arylalkylamino, aralkylamino, alkylaralkylamino, arylaralkylamino, hydroxy, alkoxy, aryloxy, aralkyloxy, mercapto, alkylthio, arylthio, aralkylthio, carboxyl, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, cyano, hydroxysulfonyl, amidosulfonyl, dialkylamidosulfonyl, arylalkylamidosulfonyl, formyl, acyl, aroyl, alkyl, alkylene, alkenyl, aryl, aralkyl, vinyl, alkynyl group and by the corresponding substituted groups; - m = 0 - 1; n = 1-4, A is nil, O, or NH; R9 represents an alkylene group; -Z is H, amino, dialkylamino, or trialkylamino salt; X- is an anion; R5, R6, R7 and R8 are independently H or C1-C6 alkyl or R1 in combination with R5 or R6, or R2 in combination with R5 or R6, or R3 in combination with R7 or R8, or R4in combination with R7 or R8 represents an alkylene, alone or in association with a pharmaceutically acceptable carrier. These compounds, which are usefull as intermediate and as bactericides, as antiviral agent and in the treatment of immunologic disoders.

Description

HALOGENATED RODAMINE DERIVATIVES AND APPLICATIONS THEREOF Field of the Invention The invention relates to novel rhodamine derivatives which are useful for their pharmaceutical and non-pharmaceutical properties. The rhodamine derivatives of the present invention exhibit powerful bactericidal and antiviral activities. They are also useful, alone or in association with a pharmaceutically acceptable carrier, in the treatment and / or prevention of immune disorders. Moreover, these derivatives are useful as intermediates in the synthesis of new additional rhodamine derivatives, and also in the new synthesis of known rhodamine derivatives. Finally, the present invention also relates to new processes for the preparation of rhodamine derivatives.

BACKGROUND OF THE INVENTION Photodynamic therapy has been used as a method for the eradication of neoplastic cells from autologous grafts for cancer treatments. This method is based on the use of photosensitizing dyes, which, when they are activated with light of a particular wavelength, produce toxic "O2" radicals, which eventually lead to cell death. Photo-chemical treatments have also been used to inactivate the pathogen, such as in the "decontamination" of blood and blood products. The danger of transmission of pathogens through the transfusion of whole blood, platelet concentrates, plasma, and / or red blood cells, still represents a major concern in medicine. Although there has been an im- pressionante progress in the prevention and maintenance of the safety of the blood with respect to the presence of microorganisms, the components of the blood continue to carry risk of transfusion of pathogens. Moreover, the presence of viruses in the blood components is also of great concern, mainly due to the presence of Hepatitis C and the human immunodeficiency virus (HIV), even when the risk of contamination is reduced to negligible levels. The presence of other viruses is also required, and includes the lin-photrophic human T-cell virus type 1 (HTLV-1), Hepatitis B (HBV), and cytomegalovirus. Photodimic compounds, such as pseuralens, porphyrins, riboflavin, and dimethylmethylene blue have been used in the treatment of pathogens in the blood product. These compounds need radiation by means of an ultraviolet A lamp (UVA) to activate, thus leading to a possible mutagenic effect in the remaining cells present in the treated samples. (Co-rash, L. Inactivation of infectious, pathogens in labile blood components: meeting the challenge, Transfus Clin Biol, 2001, 8, 138-145; Lin L., Londe, H., Janda, MJ, Hanson, CV and Corash, L., Photochemical inactivation of pathogenic bacteria in human platelet concentrates, Blood, 1994, 83, 9, 2698-2706; Lin, L, Londe, H., Hanson, CV, Wiesehahn, G., Isaacs, S. Cimino , G. and Corash, L., Photochemical inactivation of cell-associated human immunodeficiency virus in piatelet concentrates, Blood, 1993, 82, 1, 292-297; Lin, L., Eiesehalm, GP, Morel, PA and Corash , L., Use of 8-methoxypsoralen and long-wavelength ultraviolet radiation for decontamination of platelet concentrates, Blood, 1989, 74, 1, 517-525; Lin, L., Cook, DN, Wiesehahn, GP, Alfonso, R. , Behrman, B., Cimino, GD, Corten, L., Damonte, PB, Dike-man, R, Dupuis, K., Fang, YM, Hanson, CV, Heasrt, JE, Lin, CY, Londe, H. , Metchette, K., Nerio, AT, Pu, JT, Reames, AA, Rheinschmidt, M., Tessman, J., Is aacs, S.T., Wollowitz, S. and Corash, L., Photochemical inactivation of vi-ruses and bacteria in platelet concentrates by use of a novel psoralen and long-wavelength ultraviolet light, Transfusion, 1997, 37, 423-435). Due to the UVA exposure of the blood components, these techniques are not entirely satisfactory. Consequently, there was a need for new light-sensitive derivatives that would not need the ex- position to UVA of the components of the blood, and that could also be a safer and more acceptable replacement for blood components treated with UVA. Immune disorders are uncontrolled cell proliferations that result from the production of immune cells that recognize normal cells and tissues as foreign. After a period of variable latency during which they are clinically silent, cells with immunoreactivity to normal cells, induce damage in these cells and normal tissues. These immune-immunological disorders are usually divided into alloimmune conditions and autoimmune conditions. The alloimmune disorders occur primarily in the context of allogeneic transplantation (bone marrow and other organs: kidney, heart, liver, lung, etc.). In the bone marrow transplant facility, the donor immune cells present in the hematopoietic stem cell graft react to normal host tissues, causing graft-versus-host disease (GVHD). Injectable disease against the host induces damage primarily to the liver, skin, colon, lung, eyes, and mouth. Autoimmune disorders are comprised of a number of arthritic conditions, such as rheumatoid arthritis, scleroderma, and lupus erythematosus; endocrine conditions, such as diabetes elitus; neurological conditions, such such as multiple sclerosis and myasthenia gravis; hematological disorders, such as autoimmune hemolytic anemia, etcetera. The immune reaction, both in alloimmune and autoimmune disorders, progresses to generate organ dysfunction and damage. Despite important advances in treatment, immunological complications continue to be the main cause of failure of allogeneic transplants, either in hematopoietic cell transplantation (GVHD) or in solid organ transplantation (graft rejection). In addition, autoimmune disorders represent a major cause of both morbidity and mortality. The prevention and treatment of these immune disorders has relied mainly on the use of immunosuppressive agents, therapies based on monoclonal antibodies, radiation therapy, and more recently, molecular inhibitors. There has been a significant improvement in the results with the continuous development of combined modalities, but for a small number of disorders and patients. However, for the most frequent types of transplants (bone marrow, kidney, liver, heart, and lung), and for most immune disorders (rheumatoid arthritis, connective tissue diseases, multiple sclerosis, etc.), achieved the resolution of immune dysfunction and cure. As a result, the development of new approaches for the prevention and treatment of patients with immune disorders, in particular for patients who are at high risk, or whose disease has progressed and are refractory to conventional immunosuppressive therapy. Allogeneic stalk cell transplantation (AlloSCT) has been used to treat a number of malignant and non-malignant conditions. Allogeneic stalk cell transplantation is based on the administration of high-dose chemotherapy with and without total body irradiation to eliminate malignant cells and host's hematopoietic cells. The stem cells of the normal hematopoietic donor are then infused into the patient, in order to replace the host's hematopoietic system. It has been shown that transplantation of allogenic stem cells induces higher response rates, compared with conventional therapeutic options. An important issue that needs to be emphasized when allogeneic stalk cell transplantation is used is related to the risk of reinfusing immune cells that will subsequently recognize the patient's cells as foreign, and will cause graft-versus-host disease. A variety of techniques have been developed that can deplete up to 105 of the T cells from the marrow or peripheral blood. These techniques, including the immunological and pharmacological purge, are not entirely satisfactory. An important consideration when stem cell grafts are purged, T cells not reactive with the host will be retained, so that they can exert an anti-infectious and anti-leukemia activity after grafting. The potential of fo-todynamic therapy, in association with photosensitizing molecules capable of destroying immunologically reactive cells, while dispersing normal immune cells not reactive with the host, to purge hematopoietic cell grafts in preparation for transplantation of allogeneic stem cells or transplantation of autologous stem cells (AutoSct), and after transplantation of allogeneic stem cells in the context of donor lymphocyte infusions, to eliminate recurrent leukemia cells, has been unexploited in much. To achieve the eradication of T cells, several approaches have been proposed, including: 1) in vitro exposure of the graft to monoclonal antibodies and immunotoxins against the antigens present on the surface of T cells (anti-CD3, anti- CD6, anti-CD8, etc.); 2) the selection in vi tro by the soybean agglutinin and the rosette formation of sheep red blood cells; 3) the positive selection of CD34 + stem cells; Y 4) in vivo therapy with combinations of anti-thymocyte globulin, or monoclonal antibodies; 5) the in vitro exposure of the donor T cells that react with the receptor by monoclonal antibodies or immunotoxins directed to the interleukin 2 receptor or the OX-40 antigen (Cavazzana-Calvo M. et al. (1990) Transplantation, 50: 1-7; Tittle TV et al. (1997) Blood 89: 4652-58; Harris DT et al. (1999) Bone Marrow Transplantation 23: 137-44). However, most of these methods do not specifically target the subset of alloreactivity T cells, and are associated with numerous problems, including disease recurrence, graft rejection, second malignancies, and severe infections. In addition, the clinical relevance of several of these methods should be established. There are many reports on the use of photodynamic therapy in the treatment of malignancies (Daniell M. D., Hill J. S. (1991) Aust. N. Z. J. Surg., 61: 340-348). The method has been applied for cancers of different origins, and more recently for the eradication of viruses and pathogens (Raab O. (1990) Infusoria Z. Biol., 39: 524). Initial experiments on the use of photodynamic therapy for the treatment of cancer using different photoactivable substances that occur naturally or synthetically produced, were published early in century (Jesionek A., Tappeiner V.H. (1903) Muench Med Wo-chneshr, 47: 2042, Hausman W. (1911) Biochem. Z, 30: 276). In the 1940s and 1960s, a variety of tumor types underwent photodynamic therapy in vitro and in vivo (Kessel, David (1990) Photodynamic Therapy of Neoplastic Disease, Volumes I, II, CRC Press, David Kessel, Ed. ISBN 0-8493-5816-7 (v. 1), ISBN 0-8493-5817-5 (v.2)). Dougherty et al. And others, in the 1970s and 1980s, systematically explored the potential of the oncological application of photodynamic therapy (Dougherty TJ (1974) J. Nati, Cancer Inst., 51: 1333-1336, Dougherty TJ et al. (1975) J. Nati. Cancer Inst. 55: 115-121; Dougherty TJ et al. (1978) Cancer Res., 38: 2628-2635; Dougherty TJ (1984) Urol. Suppl. 23:61; Dougherty TJ (1987) Photochem, Photobiol 45: 874-889).

Treatment of Immunoreactive Cells with Photodynamic Therapy There is currently a lack of agents that allow the selective destruction of the immunoreactive cells, while leaving the normal but suppressed residual cell population intact. Previously, the preferential recovery of the photosensitive dye and the cytotoxicity of the photodynamic therapy against leukemia (Jamieson C.H. et al. (1990) Leuk, Res., 14: 209-219) and the cells have been demonstrated. lymphoid (Greinix H.T. et al., Blood (1998) 92: 3098-3104, reviewed in Zic J.A. et al, Therapeutic Apheresis (1999) 3: 50-62). It would be highly desirable to provide photosensitizers having at least one of the following characteristics: i) preferential localization and recovery by immunoreactive cells; ii) after the application of appropriate light intensities, annihilation of the cells that have accumulated and retained the photosensitizing agents; iii) dispersion of the normal hematopoietic stem cell compartment from the destructive effects of the activated photosensitizers; and iv) potential use of photosensitizers for the purging of haematopoietic stem cells from immunoreactive cells in preparation for transplantation of allogeneic or autologous stem cells; v) potential use of photosensitizers for the ex vivo elimination of reactive immune cells in patients with immune disorders.

Rhodamine Dyes Rhodamine 123 (2- (6-amino-3-imino-3H-xanten-9-yl) benzoic acid methylester hydrochloride), a thin dye. lipophilic tiónico of the class of pirilio, can alter the cellular homeostasis and can cytostatic or cytotoxic on a exhibition of high concentration and / or photodynamic therapy, although with a very poor quantum performance (Darzyn-kiewicz Z., Cárter S. (1988 ) Cancer Res., 48: 1295-1299). It has been used in vitro as a specific fluorescent dye for living mitochondria. It is recovered and is preferentially retained by many types of tumor cells, damaging their proliferation and survival by altering the function of the membrane and mitochondria (Oseroff AR (1992) In Photodynamic therapy (Henderson BW Dougherty TJ, editors) New York: Marcel Dekker, pages 79-91). In vivo, chemotherapy with rhodamine 123 may prolong the survival of the cancerous mice, but, despite initial attempts to use rhodamine 123 in the treatment of tumors, the systemic toxicity of rhodamine 123 may limit utility (Bernal, SD, et al. (1983) Science, 222: 169; Powers, SK et al. (1987) J. Neurosur., 67: 889). The Patent of the United States of North America Number 4,612,007, issued September 16, 1986 in the name of Richard L. Edelson, discloses a method for externally treating human blood, with the aim of reducing the functioning of the lymphocyte population in the blood system of a human subject . The blood, extracted from the to, is passed through a field of ultraviolet radiation in the presence of a dissolved photoactive agent able to form photoadducts with the lymphocyte DNA. This method has the following disadvantages and deficiencies. The method described is based on the use of known commercially available photoactive chemical agents for the external treatment of the patient's blood, leaving the bone marrow and leukaemic clones potential residents intact in the process. According to Richard L. Edelson, the method only reduces, and does not eradicate, the white cell population. Moreover, the wavelength range of the ultraviolet radiation used in the process proposed by Richard L. Edelson could be harmful to normal cells. The International Application published on January 7, 1993 under International Publication Number WO 93/00005, discloses a method for inactivating pathogens in the body fluid, while minimizing the adverse effects caused by the photosensitive agents. This method consists essentially in the treatment of the cells in the presence of a photoactive agent under conditions that effect the destruction of the pathogen, and in preventing the treated cells from contacting the extra extracellular protein for a predetermined period of time. This method refers to the eradication of the infectious agents of the collected blood and its components, before of its storage or transfusion. It would be highly desirable to provide novel rhodamine derivatives for the treatment of immunoreactive cells, which overcome these drawbacks, as long as they do not have systemic toxicity for the patient. Halogenated rhodamine salts are dyes that have the property of penetrating the cells and of being located generally in the mitochondria. They have been used in conjunction with photoactivation to kill certain types of cells, that is, cancer cells in leukemia, and activated T cells in autoimmune diseases. The generally accepted mechanism for the killing effect of cells is the production of oxygen from single-te, which is the reactive intermediate in the alteration of the biological processes that sustain the life of the cell. The role of rhodamine dye in the production of singlet oxygen is that of a photosensitizer, that is, of a molecule that absorbs the energy of incident light, and transfers it to oxygen in the ground state, thereby elevating it to its excited status of singlet, which is the reactive intermediary. Furthermore, it is known that the efficiency of the energy transfer process is greatly improved by the presence of heavy atoms, such as halogens, on the aromatic chromophore of the dye.

A critical problem that has not been solved, however, is the differential recovery of the photosensitizer by the target cells in relation to the other normal cells. Actually, it is known that recovery is generally a function of the molecular structure of the dye that is absorbed, and that this property varies with different cell types. Accordingly, it would be highly desirable to provide a series of novel halogenated rhodamine dyes carrying a variety of substituents at different positions in the molecule, thereby making new selective dyes available for specific target cells. An object of the present invention is to produce new photosensitizers endowed with the following characteristics: i) localization and preferential recovery by the immunoreactive cells; ii) after the application of appropriate light intensities, annihilation of these cells that have accumulated and retained the photosensitizing agents; iii) dispersion of the normal hematopoietic stem cell compartment from the destructive effects of the activated photosensitizers; iv) potential use of photosensitizers for the purging of hematopoietic stem cells from immunoreactive cells in preparation for the transplantation of allogeneic or autologous stem cells; and v) potential use of photosensitizers for the ex vivo elimination of reactive immune cells in patients with immune disorders. Accordingly, in accordance with the present invention, a series of novel rhodamine derivatives are provided alone or in association with a pharmaceutically acceptable carrier; whereby, the photoactivation of these derivatives induces cell annihilation, while the inactivated derivatives of the general structure represented by the formula (I), and the salts thereof, are substantially non-toxic to the cells. In accordance with the present invention, the use of the photoactivatable rhodamine derivatives according to the invention is also provided for the fo-todynamic treatment for the selective destruction and / or inactivation of the immunologically reactive cells, without affecting the normal cells, and without causing systemic toxicity to the patient, where the appropriate intracellular levels of these derivatives are reached, and irradiation of an appropriate wavelength and intensity is applied. In accordance with the present invention, there is also provided a method for the prevention of graft-versus-host disease associated with transplantation of alogenic stem cells in a patient, which comprises the steps of: a) activating the lymphocytes of a donor by mixing the donor cells with the host cells for a sufficient time, for a sufficient period of time for a reaction to occur immune; b) substantially removing activated lymphocytes from step a) with photodynamic therapy, using a therapeutic amount of a photoactivatable derivative or a composition of claim 1 under irradiation of a suitable wavelength; and c) performing allogeneic stem cell transplantation using the treated mixture from step d). In accordance with the present invention, there is provided a method for the treatment of an immune disorder in a patient, which comprises the steps of: a) harvesting the patient's hematopoietic cells; b) treating the hematopoietic cells ex vivo from step a) by photodynamic therapy, using a therapeutic amount of a photoactivatable derivative or a composition of claim 1 under irradiation of a suitable wavelength; and c) performing the infusion of the graft or transplanting the autograft using the hematopoietic cells treated of step b). The immunological disorder can be selected from the group consisting of conditions in which the cells themselves or the donor cells react against host tissues or foreign targets, such as graft-versus-host disease, graft rejection, disorders autoimmune, and T-cell-mediated immuno-allergies. Hematopoietic cells can be selected from the group consisting of bone marrow cells, peripheral blood cells, and spinal cord blood mononuclear cells. For the purpose of the present invention, the following terms are defined below. The term "immunoreactive disorders" is intended to mean any reaction and / or alloimmune or autoimmune. According to other aspects of the present invention, these rhodamine compounds are prepared following the general strategy of halogenating readily available rhodamine dyes, thus generating a first varied series of intermediates, which can serve themselves as potential photosensitizers, or use these halogenated rhodamines as intermediates in the synthesis of a second series of rhodamine dyes, in which one or more halogens have been replaced by one of the groups post of the structure (I). In the case where all the halogens are replaced by new groups, a subsequent halogenation step is added to the sequence to obtain the desired compound of structure I (see Figures 1 to 5) - The test of these compounds on different cell types surprisingly revealed some of the candidate molecules as non-toxic, more efficient, and more selective than the known halogenated rhodamine dyes.

COMPENDIUM OF THE INVENTION The present invention relates to rhodamine derivatives of the formula (I): © wherein: one of Ri, R2, R3, R, and Rio represents a halogen atom, and each of the remaining Ri, R2, R3, R4, and One of the remaining Rio group is independently selected from the group consisting of hydrogen, halogen atoms, an amino group, acylamino, dialkylamino, cycloalkylamino, azacycloalkyl, alkylcycloalkylamino, aroylamine, diarylamino, arylalkylamino, aralkylamino, alkylaryl- Quilamino, arylaralkylamino, hydroxy, alkoxy, aryloxy, aralkyloxy, mercapto, thioalkyl, thioaryl, thioaralkyl, carboxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, cyano, hydroxysulfonyl, amidosulfonyl, dialkylamidosulfonyl, arylalkylamidosulfonyl, formyl, acyl, aroyl, alkyl, alkylene, alkenyl, aryl, aralkyl, vinyl, alkynyl, and the corresponding substituted groups; m = 0-1; - n = 1-4; A is nothing, O, or NH; - Rg represents an alkylene group; Z is H, amino, dialkylamino, or a trialkylamino salt; - X "is an anion, and R5, Re, R7, and R8 are independently H or alkyl of 1 to 6 carbon atoms, or Ri in combination with R5 or R ?, or R2 in combination with R5 or Re, or R3 in combination with R7 or R8, or R4 in combination with R7 or R8, represents an al-kylene group, alone or in association with a pharmaceutically acceptable vehicle. The invention also relates to intermediates of the formulas (II) to (VII), and those of the formula (I '), as defined in 1 to 5, which are useful, among other things, in the synthesis of the rhodamine derivatives of the formula (I) • The invention also relates to the new processes for the synthesis of new rhodamine derivatives of the formula (I), wherein the different groups Ri to Rio, A, X, Y, Y 'and Z, y and n are as defined above, without the exclusion of the compounds listed in the condition at the end of the previous definition. These processes are defined by the schemes represented by Figures 1 to 5, and by the corresponding parts of the description. The rhodamine derivatives of the invention are useful alone or in combination with a vehicle, for the treatment of infections generated by Gram + and / or Gram- bacteria. As well as in the treatment of diseases generated by enveloped viruses or by non-enveloped viruses. These compounds are also useful in the in vivo and ex vivo treatment of immune disorders.

BRIEF DESCRIPTION OF THE SCHEMES Figure 1 is a general summary of the derivations two substituted rhodamine 4- and 2, 7-halogenated. Figure 2 is the general synthesis of substituted rhodamine 2- and 4, 5-halogenated derivatives. Figure 3 is the general synthesis of substituted rhodamine 4- and 2, 7-halogenated derivatives. Figure 4 is the general synthesis of substituted rhodamine 2- and 4, 5-halogenated derivatives. Figure 5 is the general synthesis of substituted rhodamine 2- and 4, 5-halogenated derivatives. Figure 6 is the bacteriostatic activity of rhodamine derivatives against E. coli; the bacterial strain of E. coli was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in a reduction of the bacterial growth record: HA-X-40: 0.6 log; XA-X-44: eradication; HA-X-164; 0.25 log; HA-X-171: 3.7 logs; HA-VIII-92; 6.2 logs; TH9402: 7 logs. LB is growth without compounds. Figure 7 is the bacteriostatic activity of rhodamine derivatives against P. aeruginosa} the bacteria strain of P. aeruginosa was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in the reduction of the bacterial growth record: TH9402: 2 logs. LB is growth without compounds. Figure 8 is the bacteriostatic activity of rhodamine derivatives against S. typhimurium; the bacterial strain riana de S typhimurium was treated with the 50 μM rhodamine derivatives without extrusion time. The determined effects are expressed in a reduction of the bacterial growth record: XA-A-44: 5 logs; HA-X-164; 0.3 logs; TH9402; 6.7 logs LB is growth without compounds. Figure 9 is the bacteriostatic activity of rhodamine derivatives against E. coli; the bacterial strain of E. coli was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in a reduction of the bacterial growth record: XA-X-40: 0.6 log; XA-X-44; eradication; HA-X-164; 0.25 log; HA-X-171; 3.7 logs; HA-VIII-92; 6.2 logs; TH9402: 7 logs. LB is growth without compounds. Figure 10 is the bacteriostatic activity of rhodamine derivatives against P. aeruginosa; the bacterial strain of P. aeruginosa was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in the reduction of the bacterial growth record: TH9402: 2 logs. LB is growth without compounds. Figure 11 is the bacteriostatic activity of rhodamine derivatives against S. typhimurium; the bacterial strain of S. typhimurium was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in the reduction of the bac terial growth record: XA-X-44: 5 logs; HA-X-164: 0.3 log; TH9402: 6.7 logs LB is growth without compounds. Figure 12 is the bacteriostatic activity of rhodamine derivatives against E. coli; the bacterial strain of E. coli was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in a reduction of the bacterial growth record: HA-X-40: 0.6 log; XA-X-44; eradication; HA-X-164: 0.25 log; HA-X-171: 3.7 logs; HA-VIII-92: 6.2 logs; TH9402: 7 logs. LB is growth without compounds. Figure 13 is the bacteriostatic activity of rhodamine derivatives against P. aeruginosa; the bacterial strain of P. aeruginosa was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in a reduction of the bacterial growth record: TH9402: 2 logs. LB is growth without compounds. Figure 14 is the bacteriostatic activity of rhodamine derivatives against S. typhimurium; the bacterial strain of S. typhimurium was treated with rhodamine derivatives at 50 μM without extrusion time. The determined effects are expressed in a reduction of the bacterial growth record: XA-X-44: 5 logs; HA-X-164: 0.3 log; TH9402: 6.7 logs. LB is growth without compounds. Figure 15 is the antiviral activity of the rhodamine derivatives tested on cytomegalovirus; reductions in viral infectivity and proliferation in the FS cells. The compounds were added at 50 μM without extrusion time. Reductions in the registry of viral infectivity and proliferation in FS cells; the compounds were added at 50 μM without extrusion time and without activation by light. Figure 16 is Staphylococcus epidermitis; TH9402 inhibits the bacterial growth of S. epidermitis at 50 μM without extrusion time. Figure 17 is Staphylococcus epidermiis; HA-X-40 exhibits a bacteriostatic effect on S growth. epidermitis with a reduction of two records of bacterial growth at 50 μM without extrusion time. Figure 18 is Staphylococcus epidermiis; HA-X-40 eradicates the bacterial growth of S. epidermitis at 50 μM with an extrusion time of 90 minutes. Figure 19 is Staphylococcus epidermitis; XA-X-44 eradicates the bacterial growth of S. epidermitis at 50 μM without extrusion time. Figure 20 is Staphylococcus epidermitis; HA-X-149 exhibits a bacteriostatic effect on S growth. epidermitis with a reduction of 4.5 records of bacterial growth at 50 μM without extrusion time. Figure 21 is Staphylococcus epidermitis; HA-X-164 exhibits a bacteriostatic effect on S growth. epidermitis with a reduction of 3 records of bacterial growth at 50 μM without extrusion time.

Figure 22 is Staphylococcus epidermitis; HA-X-171 exhibits a bacteriostatic effect on S growth. epidermitis with a reduction of 6.5 records of bacterial growth at 10 μM without extrusion time. Figure 23 is Staphylococcus epidermitis; HA-VIII-92 exhibits a bacteriostatic effect on S-growth. epidermitis with a reduction of 4 records of bacterial growth at 10 μM without extrusion time. Figure 24 is the antiviral activity of rhodamine derivatives tested on cytomegalovirus; Registry reductions in viral infectivity and proliferation in MRC-5 cells. The compounds were added at 50 μM without extrusion time. The following references mean: HA-X-164: the acetate salt of the 2,7-dibromo-rhodamine methyl ester B (4) HA-X-149: the hexyl ester acetate salt of 2,7-dibromo-rhodamine B (8) HA-X-171: 4, 5-dibromo-rhodamine 6G (11) HA-X-40: 2'- (6-dimethylamino-3-dimethylimino-3H-xanten-9-) methyl ester hydrochloride il) - 4 ', 5'-dichloro-benzoic acid (10) HA-X-44: 2- (2-methoxyethoxy) ethyl ester of 4,5-dibromo-rhodamine 110 (13) HA-VIII-92: 3-bromopropylester of rhodamine B (14) TH 9402: 4,5-dibromo-rhodamine 123 methyl ester.

DETAILED DESCRIPTION OF THE INVENTION A first object of the present invention is constituted by the new rhodamine derivatives of the formula (I): wherein: - one of Ri, R2, R3, R4, and Rio represents a halogen atom, and each of the remaining Ri, R2, R3, R4, and each of the remaining group Ro, is independently selected from from the group consisting of hydrogen, halogen atoms, an amino group, acylamino, dialkylamino, cycloalkylamino, azacycloalkyl, alkylcycloalkylamino, aralkylamino, diarylamino, arylalkylamino, aralkylamino, alkylarylamino, arylaralkylamino, hydroxy, alkoxy, aryloxy, aralkyloxy, mercapto , thioalkyl, thioaryl, thioaralkyl, carboxyl, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, cyano, hydroxysulfonyl, amidosulfonyl, dialkylamidosulfonyl, arylalkylamidosulfonyl, formyl, acyl, aroyl, alkyl, alkylene, alkenyl, aryl, aralkyl, vinyl, alkynyl, and the corresponding substituted groups; m = 0-1; n = 1-4; A is nothing, O, or NH; - R9 represents an alkylene group; Z is H, amino, dialkylamino, or a trialkylamino salt; X "is an anion, and R5, R ?, R, and R8 are independently H or alkyl of 1 to 6 carbon atoms, or Ri in combination with R5 or Re, or R2 in combination with R5 or Re, or R3 in combination with R7 or R8, or R4 in combination with R7 or R8, represents an alkylene group, alone or in association with a pharmaceutically acceptable carrier, with the proviso that the following specific compounds are excluded: dibromo-rhodamine 123 (2- (4,5-dibromo-6-amino-3-imino-3H-xanten-9-yl) -benzoic acid methyl ester hydrochloride) also referred to as TH9402; 4, 5-dibromo-rhodamine 123 (2- (4,5-dibromo-6-amino-3-imino-3 H-xanten-9-yl) -benzoic acid ethyl ester hydrochloride); 4, 5-dibromo-rhodamine 123 (2- (4,5-dibromo-6-amino-3-imino-3H-xanthen-9-yl) -benzoic acid octy-hydrochloride); 4,5-dibromo-rhodamine n-butylester 110 (2- (4,5-dibromo-6-amino-3-imino-3H-xanthen-9-yl) -benzoic acid n-butylester hydrochloride); and rhodamine B-n-butylester (2- (6-ethylamino-3-ethylimino-3H-xanthen-9-11) -benzoic acid n-butylester hydrochloride). According to a preferred embodiment of this object of the invention: "alkyl" means a straight or branched chain aliphatic hydrocarbon group, and the corresponding substituted alkyl group bearing one or more substituents which may be the same or different, and which selected from the group consisting of halogen, aryl, hydroxyl, al-coxyl, aryloxy, alkyloxy, thioalkyl, thioaryl, aralkyloxy, thioaralkyl, and cycloalkyl, and "branched" means that a lower alkyl group, such as methyl, ethyl, or propyl, is attached to a linear alkyl chain, and preferred alkyl groups include the "lower alkyl" groups, which are the alkyl groups having from about 1 to about 6 carbon atoms, and the alkyl groups of example are the methyl, ethyl, isopropyl, hexyl, cyclohexylmethyl groups, and methyl or ethyl groups are more preferred; "cycloalkyl" means a non-aromatic ring preferably composed of 4 to 10 carbon atoms, and the cyclic alkyl group may be partially unsaturated, and preferred cyclic alkyl rings include cyclopentyl, cyclohexyl, cyclo-octyl; the cycloalkyl group may be optionally substituted with an aryl group substituent, and the cyclopentyl and cyclohexyl groups are preferred; "alkenyl" means an alkyl group containing a carbon-carbon double bond, and preferably having from 2 to 5 carbon atoms in the straight chain, and the example groups include allylvinyl; "alkynyl" means an alkyl group containing a carbon-carbon triple bond, and preferably having from 2 to 5 carbon atoms in the straight chain, and the example groups include ethynyl and propargyl; "aryl" means an aromatic carbocyclic radical or a substituted carbocyclic radical preferably containing from 6 to 10 carbon atoms, such as phenyl or naphthyl, or phenyl or naphthyl substituted by at least one of the substituents selected from the group consti -removed by alkyl, alkenyl, alkynyl, aryl, aralkyl, hydroxyl, alkoxy, aryloxy, aralkoxy, carboxyl, aroyl, halogen, nitro, trihalomethyl, cyano, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acylamino, aralkylamino, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, thioalkyl, thioaryl, alkylene, or -NYY ', wherein Y and Y 'are independently hydrogen, alkyl, aryl, or aralkyl; - "aralkyl" means a radical wherein an aryl group is substituted by an alkyl H atom, and the example aralkyl group is benzyl; , - "acyl" means an alkyl-CO- group, where j the alkyl group is as described above, and the preferred acyl has an alkyl group containing from 1 to 3 carbon atoms in the alkyl group, and the groups examples include acetyl, propanoyl, 2-methylpropanoyl, bu-tanoyl, or palmitoyl; "Aroyl" means an aryl-CO- group, wherein the aryl group is as described above, and preferably contains from 6 to 10 carbon atoms in the aniol, and the example groups include benzoyl and 1- and 2-naphthoyl; ("alkoxy" means an alkyl-O- group, where the alkyl group is as described above, and the example alkoxy groups include methoxy, ethoxy, normal propoxy, isopropoxy, normal butoxy, and heptoxy; - "aryloxy" means an aryl-O- group, in wherein the aryl group is as described above, and exemplary aryloxy groups include phenoxy and naphthoxy-lo; "thioalkyl" means an alkyl-S- group, wherein the alkyl group is as described above, and the exemplary thioalkyl groups include thiomethyl, thioetyl, thioisopropyl, and thioheptyl; "thioaryl" means an aryl-S- group, wherein the aryl group is as described above, and the exemplary thioaryl groups include thiophenyl and thionaphthyl; "aralkyloxy" means an aralkyl-0- group, wherein the aralkyl group is as described above, and the example aralkyloxy group is benzyloxy-lo; "thioaralkyl" means an aralkyl-S- group, wherein the aralkyl group is as described above, and the example thioaralkyl group is thiobenzyl; "dialkylamino" means a group -NYY ', wherein both Y and Y' are alkyl groups as described above, and example alkylamino groups include ethylamino, dimethylamino, and diethylamino; "alkoxycarbonyl" means an alkyl-O-CO- group, wherein the alkyl group is as described above, and the example alkoxycarbonyl groups include methoxy- and ethoxycarbonyl; "aryloxycarbonyl" means an aryl-O-C0- group, wherein the aryl group is as described above, and exemplary aryloxycarbonyl groups include fe-noxy- and naphthoxycarbonyl; "aralkoxycarbonyl" means an aralkyl-0-CO- group, wherein aralkyl is as defined above, and the exemplified aralkoxycarbonyl group is benzyloxycarbonyl; - "carbamoyl" is a group H2N-CO-; "alkylcarbamoyl" is a group Y'YN-CO-, wherein one of Y and Y 'is hydrogen, and the other of Y and Y' is alkyl as defined above; "dialkylcarbamoyl" is a group Y'YN-CO-, wherein both Y and Y 'are alkyl as defined above; "acylamino" is an acyl-NH group, wherein acyl is as defined above; "aroilamino" is an aroyl-NH group, where aroyl is as defined above; "alkylene" means a straight or branched chain bivalent hydrocarbon group preferably having from 2 to 8 carbon atoms, and the alkylene group may be interrupted by one or more nitrogen atoms substituted, wherein the substituent of the nitrogen is alkyl or oxygen or sulfur atoms, and it is now preferred more than the alkylene group to have from 2 to 3 carbon atoms, and the alkylene groups of example include ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), -CH2NMe- CH2-, 0-CH2-0, or -0-CH2CH2-0-; "halogen" preferably means fluorine, chlorine, bromine, or iodine; "azacycloalkyl" preferably means a saturated carbon ring of 4 to 9 members, wherein one of the methylene groups is replaced by nitrogen; "cycloalkylamine" means a group -NYY ', wherein one of Y and Y is hydrogen, and the other of Y and Y is cycloalkyl as defined above; "alkylcycloalkylamino" means a group -NYY ', wherein one of Y and Y' is alkyl as defined above, and the other of Y and Y 'is cycloalkyl as defined above; "diarylamino" means a group -NYY ', wherein both Y and Y' are aryl groups as described above; "aralkylamino" means a group -NYY ', wherein one of Y and Y' is hydrogen, and the other of Y and Y 'is aralkyl as defined above; "arylalkylamino" means a group -NYY ', wherein one of Y and Y' is alkyl as defined above, and the other of Y and Y is aryl as defined above; "alkylalkylamino" means a group NYY ', wherein one of Y and Y' is alkyl as defined above, and the other of Y and Y 'is aralkyl as defined above; "arylalkylamino" means a group -NYY ', wherein one of Y and Y' is aryl as defined above, and the other of Y and Y 'is aralkyl as defined above; - "mercapto" is a group -SH or SR, wherein R can be any of the Ri to Rio groups defined above, -SH, mercaptoaryl, and mercaptoalkyl groups are preferred; "hydroxysulfonyl" is -S03H; - "amidosulfonyl" is -S02NH2; "dialkylsulphonyl" means a group -S02NYY ', wherein both Y and Y' are alkyl groups as described above; "arylalkylamidosulfonyl" means a group -S02NYY ', wherein one of Y and Y' is aryl as defined above, and the other of Y and Y 'is aralkyl as defined above; and - "anion" means the deprotonated form of an organic or inorganic acid, and the anion is preferably selected from hydrochlorides, hydrobromides, sulfates, nitrates, borates, phosphates, oxalates, tartrates, maleates, citrates, acetates, ascorbates, succinates, benzenesulfonates, methanesulfonates, cyclohexanesulfonates, toluenesulfonates, sulphamates, lactates, malonates, ethanesulfonates, cyclohexyl sulfamates, and kinetics. In the case where the whey derivative carries one or more acid substituents, the covered compound comprises the inner salt or any salt derived from neutralization by any of the following bases: sodium hydroxide, hydroxide potassium, calcium hydroxide, lithium hydroxide, ammonia, ethylenediamine, lysine, diethanolamine, piperazine, and the like. A preferred embodiment of the invention is constituted by the rhodamine derivatives wherein at least two of the groups Ri, R2, R, R4, and Rio represent a halogen atom which is preferably a bromide atom. More preferred are rhodamine derivatives wherein the halogen atoms are in the 2-7, 4-5, or 4'-5 position on the ring, or are at the end of the ester chain. The following specific rhodamine derivatives are particularly interesting: - 2'- (6-dimethylamino-3-dimethylimino-3H-xanten-9-yl) -4,5-dichloro-benzoic acid methyl ester hydrochloride; 2- (2-methoxyethoxy) ethyl ester of 4,5-dibromo-rhodamine; 2,7-dibromo-rhodamine B hexyl ester acetate salt; 2,7-dibromo-rhodamine B methyl ester acetate salt; 4, 5-dibromo-rhodamine 6G; and Rhodamine 3-bromopropylester B.

A second object of the present invention is constituted by the intermediates represented by the forms (II) to (VII) and (I '), the formulas being as defined in Figures 1 to 5, wherein the different groups are as defined above, without any ignorance. Intermediates are as defined in Figures 1 to 5. A third object of the present invention is constituted by new processes for the synthesis of new rhodamine derivatives of the formula (I), wherein the different Ri a Rio, A, X, Y, Y 'and Z, and m and n, are as defined above, without the exclusion of the compounds listed in the condition at the end of the previous definition. These processes are defined by the schemes represented in Figures 1 to 5, and by the corresponding parts of the description. A fourth object of the present invention is constituted by the use of at least one whey derivative as defined in the first object of the invention, without the exclusion of the compounds listed in the condition at the end of the definition of the rhodamine derivative of the formula (I), alone or in combination with a vehicle, for the treatment of infections generated by Gram + and / or Gram- bacteria. In accordance with a preferred embodiment of the present invention, the rhodamine derivatives are used for the treatment of infections generated by Staphylococcus epidermitis. Of particular interest are the use of 4,5-dibromo-rhodamine 2- (2-methoxyethoxy) ethyl ester as a bacteriostatic agent against Escherichia coli 0157: H7, and / or against Salmonella typhimurium LT2; - the use of 2,7-dibromo-rhodamine B hexyl ester acetate salt as a bacteriostatic agent against Salmonella typhimurium LT2; the use of 4, 5-dibromo-rhodamine 6G as a bacteriostatic agent against Escherichia coli: 0157: H7; - the use of rhodamine B 3-bromopropylester as a bacteriostatic agent against Escherichia coli 0157: H7; and the use of 4,5-dibromo-rhodamine methyl ester as a bacteriostatic agent against Escherichia coli 0157: H7, Salmonella typhimurium LT2, and / or Pseudomonas aerugi-nosa.

Preferably, for this therapeutic use, the rhodamine derivatives are combined with a vehicle which is a pharmaceutically acceptable carrier, and is preferably selected from the group consisting of 5 percent mannitol and / or dimethyl sulfoxide. However, any vehicle is possible, and the acceptable vehicle of preference is constituted by 5 percent mannitol: in water or in dimethyl sulfoxide. In the case of the 2,7-dibromo-rhodamine B hexyl ester acetate salt of HA-X-149, HA-X-164, the vehicle is preferably dimethyl sulfoxide. A fifth object of the present invention is constituted by a bactericidal composition for the treatment of a liquid contaminated with Gram + and / or Gram- bacteria, whose composition comprises an effective amount of at least one rhodamine derivative as defined above, without the exclusion of the compounds listed in the condition at the end of claim 1, alone or in combination with a vehicle. A sixth object of the present invention is constituted by a bactericidal solution, for the treatment of a site contaminated with Gram + and / or Gram- bacteria, whose solution comprises an effective amount of at least one rhodamine derivative of the formula (I). ) as defined above- mind, without any ignorance, alone or in combination with a vehicle, for the treatment of infections generated by Gra ™ and / or Gram-bacteria. A seventh object of the present invention is constituted by a method for the treatment of infections generated by Gramt and / or Gram-bacteria, which method comprises administering to a human or an animal that needs it, an effective amount of at least one derivative of rhodamine of the formula (I) as defined above, without any knowledge, alone or in combination with a vehicle. According to a preferred embodiment of this method, the effective amount administered comprises between 0.5 and 200 milligrams per kilogram of body weight per day. An eighth embodiment of the present invention is constituted by a medicament containing an effective amount of at least one rhodamine derivative of the formula (I) as defined above, without the exclusion of the compounds listed in the condition at the end of the definition , alone or in combination with a vehicle, for the treatment of infections generated by Gram + and / or Gram- bacteria. A tenth object of the present invention is constituted by the use of an effective amount of at least one derivative of rhodamine or a salt thereof as defined above, without any knowledge whatsoever, alone or in combination. with a vehicle, in the treatment of diseases developed by enveloped viruses or by non-enveloped viruses. Preferably, the enveloped virus is one with a double-stranded DNA, more preferably one of the Herpes viridae family. A eleventh object of the present invention is constituted by a medicament containing an effective amount of at least one rhodamine derivative or a salt thereof, as above, without the exclusion of the compounds listed at the end of the definition of rhodamine derivatives. of the formula (I), alone or in combination with a vehicle, for the treatment of viral infections. A further preferred embodiment of the present invention is the use of: - the methyl ester hydrochloride of 2'- (6-dimethylamino-3-dimethylimino-3H-xanten-9-yl) -4 ', 5'-dichloro-benzoic acid of 4, 5-dibromo-rhodamine; 2- (2-methoxyethoxy) ethyl of 4,5-dibromo-rhodamine 110 as an antiviral agent against cytomegalovirus; - the 4,5-dibromo-rhodamine methyl ester as an antiviral agent against cytomegalovirus; and the 2,7-dibromo-rhodamine B hexyl ester acetate salt, as an antiviral agent against cytomegalovirus. Another preferred embodiment of the invention is substituted by 2,7-dibromo-rhodamine B hexyl ester acetate as an antiviral agent against cytomegalovirus. The use of the hexyl ester acetate salt of 2,7-dibromo-rhodamine B as an antiviral agent against cytomega-lovirus. A twelfth object of the present invention is the use of the rhodamine derivatives of the formula (I) as defined above, without any lack of knowledge, in the treatment of immune disorders. According to a preferred embodiment, the use relates to improving the high throughput production of quantum and the generation of singlet oxygen on irradiation while maintaining a desirable differential retention of rhodamine between normal and cancer cells, these derivatives being of rhodamine of the formula (I) as defined above, without lack of knowledge. According to another embodiment, the use refers to the photodynamic therapy of cancer patients, by the destruction of human cancer cells, where appropriate intracellular levels of these derivatives are achieved, and irradiation of a wavelength is applied. and adequate intensity. According to a further preferred embodiment, the use of the invention relates to a method for photodynamic therapy of patients suffering from leukaemias, myelomas or disseminated multiple lymphomas, which comprises the steps of: a) harvesting the patient's human bone marrow; b) purging the bone marrow from step a) by photodynamic therapy, using a therapeutic amount of a photoactivatable derivative according to formula (I), without excluding the compounds listed in the condition at the end of the definition, under irradiation of a suitable wavelength; and c) performing autologous stem cell transplantation using the purged bone marrow from step b). Preferably, the purging of step b) further comprises intense chemotherapy and total body irradiation (TBI) procedures. Another preferred embodiment relates to a method for the in vitro purging of human bone marrow containing solid tumor metastases, selected from the group consisting of breast, lung, prostate, pancreatic metastasis, and of colonic carcinomas, melanomas and disseminated sarcomas, where surgical separation and volume reduction can be achieved, which comprises the steps of: a) harvesting the patient's human bone marrow; b) purging the bone marrow from step a) by photodynamic therapy, using a therapeutic amount of at least one photoactivatable derivative of formula (I) as it is defined above, without any lack of knowledge, under irradiation of a suitable wavelength; and c) performing autologous stem cell transplantation using the purged bone marrow from step b). Preferably, the purging of step b) further comprises intense chemotherapy and total body irradiation (TBI) procedures. A further embodiment of this object of the invention is a method for the photodynamic therapy of cancer patients, which comprises administering to these patients a therapeutically acceptable intracellular level of at least one photoactivatable derivative of the formula (I) as defined above, without any knowledge, and subject these patients to irradiation of a therapeutic wavelength properly. Preferably, at least one photoactivatable derivative is administered by instillation, injection, diffusion into the bloodstream at the tumor sites directly accessible to light emission, or at the tumor sites accessible to the laser beams, using rigid endoscopes or flexible More preferably, the laser-accessible tumor site is selected from the group consisting of urinary bladder, oral cavity, esophagus, stomach, lower digestive tract, upper respiratory tract and ferior. Another preferred embodiment of this object of the invention is constituted by a method for the treatment of leukemias in patients, which comprises the steps of: a) purging the cancerous clones from the bone marrow of the patients; b) subjecting the purged clones of step a) to a photodynamic treatment using a therapeutic amount of at least one of the photoactivatable derivatives of the formula (I) as defined above, without the lack of knowledge present at the end of the definition, under irradiation of a suitable wavelength, for the selective destruction of the leukemic cells, without affecting the normal cells of the patients; and c) administering the treated clones of step b) to patients; without causing in this way a systemic toxicity for patients. A fourteenth object of the present invention is constituted by a photoactivatable pharmaceutical composition for the selective destruction and / or inactivation of immunologically reactive cells, without affecting the normal cells, and without causing systemic toxicity for the patient, this composition comprising at least a photoactivatable rhodamine derivative of the formula (I) as defined above above, without the exclusion of the compounds listed in the condition at the end of the definition, and photoactive derivatives thereof; in association with a pharmaceutically acceptable vehicle; wherein the photoactivation of these derivatives induces the annihilation of the cells, while the inactivated derivatives are substantially non-toxic for the cells. A fifteenth object of the present invention is constituted by the use of the photoactivatable derivatives of claim 1, for the photodynamic treatment for the selective destruction and / or inactivation of the immunologically reactive cells, without affecting the normal cells, and without cause systemic toxicity for the patient, where appropriate intracellular levels of these derivatives are reached, and irradiation of an appropriate wavelength and intensity is applied. A preferred embodiment is constituted by a method of preventing graft-versus-host disease associated with allograft stem cell transplantation in a patient, which comprises the steps of: a) activating lymphocytes from a donor, mixing the donor cells with the host cells for a sufficient time for a sufficient period of time for an immune reaction to occur; b) substantially eliminate active lymphocytes Step (a) with photodynamic therapy, using a therapeutic amount of a photoactivatable composition of claim 24, under irradiation of a suitable wavelength; and c) carrying out the transplantation of allogeneic stem cells using the treated mixture of step b). Another preferred embodiment is constituted by a method for the treatment of an immune disorder in a patient, which comprises the steps of: a) harvesting the patient's hematopoietic cells; b) treating the hematopoietic cells ex vivo from step a) by photodynamic therapy, using a therapeutic amount of a photoactivatable composition of claim 24, under irradiation of a suitable wavelength; and c) performing the graft infusion or autograft transplant, using the treated hematopoietic cells of step b). Preferably, the immunological disorder is selected from the group consisting of conditions wherein the self cells or donor cells react against host tissues or foreign targets, such as graft-versus-host disease, rejection of jerto, autoimmune disorders, and immunoallergies mediated by T cells. Most preferably, hematopoietic cells are selected from the group consisting of bone marrow cells, peripheral blood cells, and spinal cord mononuclear cells. The compounds of structure I exhibit better properties such as: marker dyes for deoxynucleotides, dideoxynucleotides, and polynucleotides; novel dyes suitable for recording fluids for the inkjet process; novel dyes for fiberglass and paper; novel dyes for the eradication of infectious biological contaminants in body tissues; novel dyes applicable in photographic processes; novel dyes applicable in cancer chemotherapy; novel dyes applicable as inhibitors of the herpes simplex virus thymidine kinase, and in the treatment and / or prophylaxis of infections caused by the herpes simplex virus; novel dyes for use as optical amplifiers of polymers and laser devices; novel dyes applicable in cellular biology; novel dyes applicable in the addition of siliceous materials to give solid dye laser devices; novel pigments applicable for paints, inks, and plastics; novel organic reagents in solvent extraction of metal ions; innovative dyes applicable in the formation of new products conjugated with other dyes; novel dyes for the manufacture of optical memory discs of the CD-ROM type; novel dyes applicable in the peptide fluorophore labeling; novel dyes applicable in flow cytometric analysis; non-veined dyes applicable as spots for the detection of Mycobacterium tuberculosis by fluorescence microscopy; novel dyes applicable in the fluorescent mapping of binding sites for substrates, ligands, and inhibitors; novel dyes to study transport through the blood-brain barrier; novel dyes to study biofilm disinfection; novel dyes applicable as fluorescent probes in cellular biology; novel dyes to be used as water tracing; novel dyes for the visualization of peptide receptors by intensified fluorescence microscopy in images; novel dyes for the formation of metal chelates in analytical chemistry; novel fluorescent dyes applicable in diagnostic therapy.

Chemical synthesis These compounds are prepared following the general halogenation strategy of the known and readily available rhodamine dyes, thus generating a first series of intermediaries, which themselves can serve as potential photosensitizers, or the of these halogenated rhodamines as intermediates in the synthesis of a second series of rhodamine dyes, in which one or more halogens have been replaced by one of the groups of structure I. In the case where all the halogens are replaced by new groups, a subsequent halogenation step is added to the sequence to obtain the desired compound of structure I (see illustrative schemes 1 and 2). Due to the specific retention of rhodamine 123 dye class by abnormal malignant cells, and the concomitant lack of accumulation by normal hematopoietic stem cells, these results provide evidence for the potential use of these three new dyes for photodynamic therapy in vivo or in vitro. In accordance with the present invention, the use of these aforementioned dyes is provided in conjugation with tumor-specific antibodies, or poisonous substances, or liposomal or lipoprotein adducts, or fluorochrome. In addition, the photosensitizers to be described have the potential to act synergistically in conjunction with other photoactive substances. Moreover, the negative selection procedure provided by the use of photodynamic treatment, does not preclude the use of other means to enrich haematopoietic stem cells, such as positive selection with anti-CD34 monoclonal antibodies.

Other Clinical Applications In addition to the use of photosensitizers in the context of in vitro bone marrow purging for metastatic leukemias and cancers, the molecules can also be used in vivo for tumor sites directly accessible to exposure to a source of light, and at appropriate local concentrations of the drugs to be described. The molecules of the invention can also be used in the photodynamic therapy of a patient suffering from disseminated multiple myelomas or multiple lymphomas. Metastatic cancers for which the therapy of this invention is appropriate include breast, lung, prostate, pancreatic, and colonic carcinomas, disseminated melanomas, and sarcomas. The photoactivatable derivatives of the present invention can be administered by instillation, injection, diffusion into the bloodstream at the tumor sites directly accessible to the emission of light from the tumor sites accessible to the laser beams using rigid or flexible endoscopes.

DESCRIPTION OF THE PREFERRED MODALITIES As a matter of illustration only, five methods of treating immune disorders involving the rhodamine derivatives according to the invention are illustrated below.

METHOD I OF TREATMENT OF LEUKEMIAS 1. Diagnostic Procedures The diagnosis of chronic myelogenous leukemia (CML) will be established using one or more of the following procedures in the blood or in the cells of the bone marrow: a) conventional cytogenetic studies with identification of Ph + metaphases that house t (9): 22); b) Fluorescent in situ hybridization for detection of reconfiguration ber / abl; and c) Southern blot analysis for the detection of a reconfigured ber fragment, or RT-PCR for the detection of chimeric ber / abl messenger RNA. 2. Bone Marrow Harvest After diagnosis, hematopoietic stem cells derived from bone marrow (DM) or peripheral blood (PB) will be harvested, using the procedures described above for autologous marrow transplantation in cancer therapy (reviewed by Herzig GP, (1981) Prog.

Hematol. , 12: 1). The hematopoietic stem cells harvested for the autograft will be treated immediately ex vivo as described below. 3. Leukemia Purge ± n Vi tro The ex vivo treatment consists of a short-term incubation of stem cells of bone marrow or peripheral blood with one or more of the selected photoactive compounds. The duration of the incubation, the cell concentration, and the molarity of the drug will be determined for each patient using an aliquot of the harvested cell population. Excess dyes will be removed by washing the cells with a medium free of sterile dye supplemented with 2 percent autologous serum. The cells are then exposed to radiant energy of sufficient intensity to effect the photodynamic purge of the leukemia cells. The effectiveness of the photodynamic purging process is verified on an aliquot of the treated cell population, before cryopreservation and / or reinfusion to the patient. Until the patient is reinfused, the cells are cryopreserved in 10 percent dimethyl sulfoxide (DMSO) - 90 percent autologous serum medium at -196 ° C in the vapor phase of liquid nitrogen. 4. Systemic Treatment of Patients Following harvesting of stem cells, the patient will be treated with conventional regimens until Indicate the autograft clinically, or immediately undergo intensive chemotherapy in doses and total irradiation of the body where indicated. 5. Autologous Stem Cell Transplantation Following appropriate treatment of the patient by high-dose chemotherapy and irradiation, and at the appropriate clinical time, the cryopreserved medullary or peripheral blood stem cells will be rapidly thawed, and diluted in a medium containing 25 IU of mi-1 DNAse, to minimize the formation of lumps. A minimum of 2x107 kilogram of nucleated cells with a viability of 85 percent to 95 percent will be returned, measured by Trypan ™ blue exclusion to the patient.

METHOD II OF TREATMENT OF MALIGNITIES 1. Diagnostic Procedures The diagnosis of malignancies will be established using conventional histopathological examination of the primary tumor. The detection of marrow involvement by neoplastic cells will be achieved by direct histological examination and ancillary procedures where indicated (ie, immuno-peroxidase, immunohistochemical, tumor marker, and hybridization studies). 2. Harvest of Bone Marrow After diagnosis, cells will be harvested hematopoietic stem cells derived from bone marrow (DM) or peripheral blood (PB), using the procedures described above for autologous marrow transplantation in cancer therapy (reviewed by Herzig GP, (1981) Prog. Hematol., 12: 1) . The hematopoietic stem cells harvested for the autograft will be treated immediately ex vivo as described below. 3. Purging Leukemia ± n v ± tro The ex vivo treatment will consist of a short-term incubation of stem cells of bone marrow or peripheral blood with one or more of the selected photoactive compounds. The duration of the incubation, the cell concentration, and the molarity of the drug will be determined for each patient using an aliquot of the harvested cell population. Excess dyes will be removed by washing the cells with a medium free of sterile dye supplemented with 2 percent autologous serum. The cells will then be exposed to radiant energy of sufficient intensity to effect the photodynamic purge of the leukemia cells. Whenever a sensitive molecular marker is available, an aliquot of the treated cell population will be tested for the detection of residual neoplastic cells prior to cryopreservation and / or reinfusion to the patient. Cells will be cryopreserved in 10 percent dimethyl sulfoxide (DMSO) - 90 percent autologous serum cent, at -196 ° C in the vapor phase of liquid nitrogen. 4. Systemic Treatment of Patients Following harvesting of stem cells, the patient will be treated with conventional regimens until the autograft is clinically indicated, or will undergo immediate intensive chemotherapy in doses and total irradiation of the body where indicated. 5. Autologous Stem Cell Transplantation Following high-dose chemotherapy and irradiation, cryopreserved medullary stem cells or peripheral blood cells will thaw rapidly and be diluted in a medium containing 25 IU of DNAse ml- 1, to minimize the formation of lumps. A minimum of 2x107 kilogram of nucleated cells with a viability of 85 percent to 95 percent will be returned, measured by Trypan ™ blue exclusion to the patient.

METHOD III PREVENTION OF GRAFT DISEASE AGAINST THE GUESTS IN THE CONTEXT OF ALLOGENIC STEM CELL TRANSPLANTATION 1. Diagnosis and Identification of Immunological Differences between the Donor and the Recipient, and Graft Disease against the Host: The Transplant of allogeneic stem cells is performed for numerous neoplastic and non-neoplastic conditions ticas The hematological malignancies are comprised of leukemia, lymphoma, multiple myeloma, myelodysplastic syndromes, etcetera; and non-hematological malignancies: aplastic anemia, congenital disorders, severe immunodeficiency syndromes, rheumatoid arthritis, scleroderma, lupus erythematosus, multiple sclerosis, and other immune disorders. Graft versus host disease is a complication of allogeneic stem cell transplantation, where donor cells react against host cells, damaging target tissues (usually skin, liver, intestine, lung, lacrimal glands or salivary, etc.). The diagnosis is supported by several clinical and laboratory parameters, which are extensively reviewed in Graf t-vs. -Host Disease, Ferrara JLM, Deeg HJ, Bura-koff SJ eds, Marcel Dekker, New York, 1997. Graft versus host disease develops against the antigens present in the recipient cells, but not in the donor cells. Immunological differences between the donor and the recipient could be present at the level of major histocompatibility antigens, minor histocompatibility antigens, or antigens associated with tumors. The disparity will be established using one or more of the following procedures on blood or bone marrow cells: a) HLA typing: serological typing conventional or molecular to identify disparities between donor and recipient in the major complex histocompatibility class I and class II antigens; and b) culture of mixed lymphocytes to identify differences in class II antigens; and c) minor histocompatibility antigens: although a few cytotoxic T cell lines are available, and could be used to identify minor histocompatibility antigens, currently, these tests are only available for research purposes. 2. Harvest of Progenitor Cells After diagnosis, hematopoietic stem cells derived from bone marrow (BM) or from peripheral blood (PB) or blood from the spinal cord of the donor will be harvested, using the procedures described above for the transplant of allogenic progenitor cells (reviewed in Bone Marrow Transplantation, Forman SJ, Blume KG, Thomas ED eds, Blackwell Scientific Publications, Cam-bridge MA, USA, 1994). Haematopoietic stem cells from the donor harvested for the allograft will be immediately incubated with irradiated host mononuclear cells (25Gy) or other cells. Host cells mixed with the donor cells are incubated in a sterile-free medium supplemented with 20% autologous serum. and interleukin-2 for 2 days. This procedure causes the alloreactivity of the donor cells to the host, and subsequently the cell graft undergoes an ex vivo photodynamic treatment as described below. 3. Selective Purge ± n of Immunoreactive Cells Ex vivo treatment will consist of a short-term incubation of previously activated bone marrow or peripheral blood stem cells with one or more of the selected photoactive compounds. The duration of the incubation, the cellular concentration, and the olarity of the drug will be determined for each patient using an aliquot of the harvested cell population. Excess dyes will be removed by washing the cells with a medium free of sterile dye supplemented with 2 percent autologous serum. The cells will then be exposed to radiant energy of sufficient intensity to effect the photodynamic purge of the leukemia cells. The effectiveness of the photodynamic purge procedure will be verified on an aliquot of the treated cell population, before cryopreservation and / or reinfusion to the patient. Until the patient is reinfused, the cells will be cryopreserved in 10 percent dimethyl sulfoxide (DMSO) - 90 percent autologous serum medium at -196 ° C in the vapor phase of liquid nitrogen. 4. Systemic Treatment of Patients Following the harvest of stem cells, the The patient will undergo intensive chemotherapy in the dose and / or irradiation when indicated. 5. Allogeneic Stem Cell Transplantation Following the appropriate treatment of the patient by high-dose chemotherapy and / or irradiation, and at the appropriate clinical time, the stem cells of spinal cord or peripheral blood or spinal cord blood cryoprecipitate. stored will be thawed rapidly and returned to the patient.

METHOD IV TREATMENT OF GRAFT DISEASE AGAINST THE GUEST AND AUTOIMMUNE DISEASE 1. Diagnostic Procedures The diagnosis of graft-versus-host disease or immunoreactive disorders will be established using a conventional clinical, biochemical, and / or histopathological examination of the blood or the appropriate tissues. The diagnostic and predictive characteristics of graft-versus-host disease are reviewed in Graft-vs. -Host Disease, Ferrara JLM, Deeg HJ, Burakoff SJ eds, Marcel Dekker, New York, 1997. 2. Harvest of Peripheral Blood Cells After diagnosis of a severe graft-versus-host disease, of an autoimmune or immunoreactive disorder, the mononuclear cells will be harvested of peripheral blood (PB) using previously described or similar leu-coforesis procedures (reviewed in Bone Marrow Transplantation, Forman SJ, Blume KG, Thomas ED eds, Blackwell Scientific Publications, Cambridge MA, USA, 1994). The collected peripheral blood mononuclear cells of the patient will be treated immediately ex vivo as described below. 3. Elimination of the cells that mediate graft disease against the host Ex vivo treatment will consist of a short-term incubation of peripheral blood stem cells with one or more of the selected photoactive compounds. The duration of the incubation, the cell concentration, and the molarity of the drug will be determined for each patient using an aliquot of the harvested cell population. Excess dyes will be removed by washing the cells with a medium free of sterile dye supplemented with 2 percent autologous serum. The cells will then be exposed to radiant energy of sufficient intensity to effect the photodynamic purge of the activated cells, which mediates the disease of the graft against the host. 4. Administration of the Cells Photodynamically Treated to the Patients The cells passed through leukophoresis that are treated photodynamically, will be reinfused in the patient. This plan- This will make possible the elimination of a large number of circulating activated lymphocytes and other cells involved in graft-versus-host disease. In addition, the cells dispersed by the photodynamic treatment are inactivated, and their reinfusion to the patient can help restore the normal immune balance.

METHOD V OF TREATMENT OF IMMUNOLOGICAL DISORDERS 1. Diagnostic Procedures The diagnosis of autoimmune disorders will be established using a conventional clinical, biochemical, and / or histopathological examination of the blood or appropriate tissues. Severe autoimmune diseases are susceptible to autologous transplantation (reviewed in Sullivan KM and collaborators, Am. Soc. Hematol, Educ. Program Book, 1998: 198-214). 2. Harvest of Hematopoietic Stem Cells After diagnosis, mononuclear cells from bone marrow (BM), peripheral blood (PB), or spinal cord blood (CB) will be harvested, using the procedures described above for the autologous marrow transplantation in cancer therapy (reviewed in Bone Marrow Transplantation, Forman SJ, Blume KG, Thomas ED eds, Blackwell Scientific Publications, Cambridge MA, USA, 1994). The hematopoietic stem cells of the patient, harvested for the autograft, will be treated immediately ex vivo as described below. 3. In vitro elimination of the cells that mediate autoimmune disorders The ex vivo treatment will consist of a short-term incubation of stem cells of bone marrow or peripheral blood with one or more of the selected photoactive compounds. The duration of incubation, cell concentration, and molarity of the drug will be determined for each patient using an aliquot of the harvested cell population. Excess dyes will be removed by washing the cells with a medium free of sterile dye supplemented with 2 percent autologous serum. The cells will then be exposed to radiant energy of sufficient intensity to perform the photodynamic purge of the immunoreactive cells, which mediate the immune disorder. 4. Administration of Photodynamically Treated Cells to Patients Stem cell cells that are treated photodynamically will be stored (frozen or in culture). This approach will make it possible to eliminate a large number of activated lymphocytes and other cells involved in the immune disorder. In addition, the cells dispersed by the photodynamic treatment are inactivated, and their reinfusion can help restore the normal immune balance.

Following harvesting of stem cells, the patient will be treated with conventional regimens until the autograft is clinically indicated, or they will be immediately subjected to intense chemotherapy in the dose and total irradiation of the body where indicated. 5. Autologous Stem Cell Transplantation Following high-dose chemotherapy and irradiation, the cryopreserved medullary or peripheral blood stem cells will be rapidly thawed and infused into the patient. The preparation of these rhodamine derivatives of the formula (I), as defined above, without the proviso, will be more readily understood by reference to the following examples, which are given for illustrative purposes. I. Synthesis of the 2,7-dibromo-rhodamine B (4) 1-1 ptetylester acetate salt. Preparation of rhodamine methyl ester B (1) To a stirred mixture of 1.63 grams (3.40 milligrams) of rhodamine B, and 100 milliliters of methanol, hydrochloric acid was bubbled through the solution for 45 minutes, and the reaction mixture was refluxed overnight. The methanol was evaporated under reduced pressure, and the dark red residue was then purified by evaporation chromatography using a mixture of methanol and dichloromethane (1: 9) as eluent, to provide the desired product as a deep red viscous residue (1.54 grams) . Rf: 0.52 (MeOH: CH2C12 1.5: 8.5) Yield: 92 percent. Ms (FAB): Calculated for C29H33O3N2: (M-C1) +: 457.2491 Found (M-Cl) +: 457.2494 UV (MeOH):? Max 555 nm. 1-2. Preparation of dihydro-rhodamine methyl ester B (2) The rhodamine B methyl ester, 1.73 grams (3.50 mi-limols) was dissolved in 250 milliliters of dichloromethane and 100 milliliters of water. An excess of NaBH4 (solid) was added in portions with vigorous stirring, for 30 minutes, until the initial dark red color was discharged. The pale orange organic phase was separated, and the aqueous phase was extracted twice with dichloromethane. The combined organic layers were dried over Na 2 SO 4 / filtered, and evaporated under reduced pressure, and the residue was purified by evaporation chromatography using ethyl acetate as the eluting solvent. The fractions containing the product were combined, and the solvent was evaporated to give the product 2 as a rose oil (1.50 grams). Rf: 0.84 (AcOEt) Yield: 93.7 percent. 1-3: Bromination of dihydro-rhodamine methyl ester B (2) In a 250 milliliter round bottom flask, dihydro-rhodamine methyl ester B (2), 1.34 grams (2.92 mmol), and 112 milliliters of grade-spectrum methanol were introduced. The mixture was stirred at room temperature until that all the ester dissolved. 2 equivalents of propylene oxide (409 microliters, 5.85 millimoles) were added, followed by the dropwise addition of 2 equivalents of bromine (300 microliters, 5.85 millimoles). The stirring was continued at room temperature for 1 hour and 30 minutes. The volatile solvent was evaporated under reduced pressure, and the red oily residue was subjected to purification by evaporation chromatography using ethyl acetate and hexanes (0.5: 9.5) as eluent, to give the desired compound 3 as a white foam solid (570 milligrams). Rf: 0.41 (AcOEt: Hexanes 0.5: 9.5) Yield: 31.6 percent. NMR: (CD3OD) d 7.86 (dd, J = 1.44 and 7.8 Hz, 1H); 7.44 (, 1H); 7.32 (m, ÍH); 7.16 (s, 2H); 7.10 (dd, J = 1.45 and 7.8 Hz, 1H); 6.93 (s, 2H); 6.17 (s, ÍH); 3.94 (s, 3H); 3.09 (q, J = 7.09 Hz, 8H); 1.04 (t, J = 7.09 Hz, 12H). Ms (FAB): (MH) + 615.1. 1-4. Oxidation of 2,7-dibromodihydro-rhodamine B methyl ester and formation of 2,7-dibromo-rhodamine B methyl ester acetate salt (4) To a stirred solution of 2,7-dibromodihydro-rhodamine B methyl ester (3_), 400 milligrams (0.64 millimoles) in 10 milliliters of dichloromethane, was added chloranil (1.2 equivalents, 0.77 millimoles, 192 milligrams). The reaction mixture was stirred at room temperature overnight, then the reaction was stopped and the solvent was evaporated under reduced pressure to give a purple residue. The oxidized compound obtained in the previous step was dissolved in 15 milliliters of dichloromethane, and acetic acid (0.8 milli-liters) was added dropwise. The obtained clear red solution was stirred for 5 minutes, at room temperature, followed by evaporation of the volatile solvent under reduced pressure to give a purple viscous residue. The residue was purified by chromatography by evaporation using 10 percent methanol in dichloromethane as eluent, to give the desired compound 4_ as a viscous purple solid (200 milligrams). Rf: 0.29 (MeOH: CH2C12 1: 9) Yield: 45.7 percent. NMR: (CD3OD) d 8.48 (dd, J = 1.45 and 7.5 Hz, ÍH); 7.95 (m, 2H); 7.52 (dd, J = 1.6 and 7.2 Hz, ÍH); 7.45 (s, 2H); 7.38 (s, 2H); 3.79 (q, J = 8 Hz, 8H); 3.71 (s, 3H); 1.99 (s, 3H); 1.37 (t, J = 7.02 Hz, 2H). Ms (FAB): Calculated for C29H32? 3N2Br2 (MH-AcO) +: 614.0779. Found: 614.0765 UV (MeOH)? Max 577 n.

EXAMPLE II II. Synthesis of the hexyl ester acetate salt of 2,7-dibromo-rhodamine B (8) II-1. Preparation of rhodamine hexyl ester B (5) To a stirred mixture of 2.39 grams (4.98 millimoles) of rhodamine B, and 120 milliliters of 1-hexanol, hydrochloric acid was bubbled through the solution for 45 minutes, and the reaction mixture was refluxed overnight. . The 1-hexanol was then distilled off under reduced pressure, and the dark red residue was purified by evaporation chromatography using a mixture of methanol and dichloromethane (1: 9) as eluent. After the evaporation of the volatile solvents, we obtained a viscous red-green residue (2.62 grams). Rf: 0.45 (MeOH: CH2C12 1.2: 8.8). Yield: 93.5 percent.

Ms (FAB): Calculated C34H43O3N2 (M-C1) +: 527. 3273 Found: 527. 3261 UV (MeOH):? Max 555 n.

II-2. Preparation of the dihydro-rhodamine hexyl ester B (6) The rhodamine hexyl ester B (5_), 940 milligrams (1.66 millimoles) was dissolved in 200 milliliters of dichloromethane and 150 milliliters of water. An excess of NaBH (solid) was added in portions with vigorous stirring, for 30 minutes, until the initial dark red color was discharged. The pale orange organic phase was separated, and the aqueous phase was extracted twice with dichloromethane. The combined organic layers were dried over Na 2 SO 4, filtered, and evaporated under reduced pressure. The crude oily residue was purified by evaporation chromatography using ethyl acetate as eluent, yielding 794 milligrams of 6 as a pink oil.

Rf: 0.92 (AcOEt) Yield: 90 percent, II-3. Bromination of dihydro-rhodamine hexyl ester B (6) In a 100 milliliter round bottom flask, we introduced dihydro-rhodamine hexyl ester B (6), 784 milligrams (1.48 ilimoles), and 25 milliliters of grade-spectrum methanol. The mixture was stirred at room temperature until all of the ester dissolved. 2 equivalents of propylene oxide (208 microliters, 2.96 milliliters) were added, followed by the dropwise addition of 2 equivalents of bromine (152 microliters, 2.96 millimoles). Stirring was continued at room temperature for 1 hour and 30 minutes. The volatile solvent was evaporated under reduced pressure, and the red oily residue was subjected to purification by evaporation chromatography using ethyl acetate and hexanes (0.25: 9.75) as eluent, to provide 207 milligrams of the pure compound as a white foamy solid, and 123 milligrams of the impure product.

Rf: 0.61 (AcOEt: Hexanes 0.5: 9.5) Yield: 20.5 percent.

II-4. Oxidation of 2,7-dibromodihydro-rhodamine B hexyl ester and formation of 2,7-dibromo-rhodamine B hexyl ester acetate salt (8) To a stirred solution of 207 milligrams of 2, 7-dibromodihydro-rhodamine B hexy-ester (0.30 millimoles) in 8 milliliters of dichloromethane, chloranil (1.2 equivalents, 0.36 millimoles, 89 milligrams) was added. The reaction mixture was stirred at room temperature overnight, then the reaction was stopped, and the solvent was evaporated under reduced pressure to give a purple residue. The oxidized compound obtained in the previous step was dissolved in 8 milliliters of dichloromethane, and acetic acid (0.8 milliliters) was added dropwise. The obtained clear red solution was stirred for 5 minutes at room temperature, followed by evaporation of the volatile solvent under reduced pressure to give a purple viscous residue, which was purified by evaporation chromatography using 10 percent methanol in dichloromethane as eluent, to give the desired compound 8_ as a viscous purple solid (198 milligrams). Rf: 0.47 (MeOH: CH2C12 1: 9). Yield: 86.9 percent. NMR: (CD3OD) d 8.29 (dd, J = 1.5 and 7.6 Hz, ÍH); 7.82 (m, 2H); 7.40 (dd, J = 1.6 and 7.2 Hz, ÍH); 7.37 (s, 2H); 7.28 (s, 2H); 3.96 (t, J = 7.2 Hz, 2H); 3.72 (q, J = 7.05 Hz, 8H); 1.91 (s, 3H); 1.29 (t, J = 7.06 Hz, 12H); 1.08 (m, 4H); 0.79 (t, J = 7.04 Hz, 3H). Ms (FAB): Calculated for C34H42? 3N2Br2 (MH-AcO) +: 684. 1561 Found: 684. 1587 UV (MeOH):? Max 582 nm.

EXAMPLE III III. Synthesis of 2 '- (6-dimethylamino-3-dimethylimino-3H-xanten-9-yl) -4', 5'-dichloro-benzoic acid methyl ester hydrochloride (10) III-l. Preparation of 2 '- (6-dimethylamino-3-dimethylimino-3H-xanten-9-yl) -4', 5'-dichloro-benzoic acid hydrochloride (9) A mixture of 3.00 grams (21.8 millimoles) of 3- (dimethylamino) phenol, 3.00 grams (13.8 millimoles) of 4,5-dichlorophthalic anhydride, and 1.72 grams of zinc chloride, is heated in an oil bath to 165 ° C-170 ° C for 5 hours and 30 minutes with agitation. The melt is cooled and converted to powder to give a red solid. The solid is washed with hot water, it is triturated with 10 percent sodium hydroxide, and diluted with water. The separating gum is collected, washed with more sodium hydroxide and water. The resulting dye base is then triturated with concentrated hydrochloric acid. Then water is added, and the red precipitate obtained is collected and dried. The dye is then dissolved in meta-nol, and precipitated with diethyl ether, to give the _9 as a red solid (3.27 grams). Rf: 0.48 (MeOH: CH2C12 2: 8). Yield: 48 percent. NMR: (CD3OD) d 8.47 (s, ÍH); 7.72 (s, 1H); 7.22 (d, J = 9.47 Hz, 2H); 7.11 (m, 2H); 7.01 (d, J = 2.4 Hz, 2H); 3.32 (s, 12H). Ms (FAB): Calculated for C24H2i03N2Cl2 (M-C1) +: 455. 0929 Found: 455. 0938 UV (MeOH):? Max 511 nm.

III-2. Preparation of acid methyl ester hydrochloride 2 '- (6-dimethylamino-3-dimethylimino-3H-xanten-9-yl) -4', 5'-dichloro-benzoic acid (10) To a 250 milliliter round bottom flask, equipped with a magnetic stirrer, was added 738 milligrams (1.50 millimoles) of acid 9, and 40 milliliters of anhydrous dichloromethane, and 10 milliliters of anhydrous DMF. The mixture was stirred under nitrogen until all the acid was dissolved. Then an amount of 309 milligrams (1.50 millimoles) of 1,3-dicyclohexylcarbodi-imide (DCC) was added, followed by 200 microliters of methanol and 18 milligrams of 4-N, N-dimethylaminopyridine (DMAP). The mixture was stirred at room temperature overnight. Then the solvent was distilled under reduced pressure to give a red residue, which was purified by evaporation chromatography using MeOH: CH2Cl2 (1.2: 8.8) as eluent, to give 1_0 as a reddish brown solid (350 milligrams). Rf: 0.52 (MeOH: CH2CL2 2: 8). Performance: 46 percent. NMR: (CD3OD) d 8.50 (s, ÍH); 7.80 (s, ÍH); 7.18 (d, J = 9.2 Hz, 2H); 7.12 (m, 2H); 7.04 (d, J = 2.31 Hz, 2H); 3.80 (s, 3H); 3.35 (s, 12H). Ms (FAB): Calculated for C25H2303N2C12 (M-C1) +: 469. 1085 Found: 469 1078 UV (MeOH):? Max 555 nm.

EXAMPLE IV IV. Preparation of 4, 5-dibromo-rhodamine 6G (11) To a quantity of 600 milligrams (1.25 millimoles) of rhodamine 6G, dissolved in 50 milliliters of methanol, a solution of 128 was added dropwise at room temperature. microliters (2 equivalents, 2.50 millimoles) of bromine. A precipitate formed 10 minutes after the bromine addition. The mixture was stirred for 3 hours, and the solvent was evaporated under reduced pressure to give a red solid. The crude was recrystallized from methanol: diethylether (80 ml: 400 ml) to give the product 17L as a reddish green solid (585 milligrams). Rf: 0.26 (MeOH: CH2C12 1: 9). Performance: 68.5 percent. NMR (CD3OD) d 8.36 (dd, J = 1.13 and 7.44 Hz, ÍH); 7.89 (m, 2H); 7.47 (dd J = 1.46 and 6.76 Hz, ÍH); 6.98 (s, 2H); 4.07 (m, 6H); 2.29 (s, 6H); 1.28 (t, J = 7.04 Hz, 6H); 1.02 (t, J = 7.05 Hz, 3H). Ms (FAB): Calculated C28H3o03N2Br2 (MH-Br) +: 600.0623. Found: 600.0605 UV (MeOH):? Max 546 n. EXAMPLE V V. Synthesis of 4,5-dibromo-rhodamine 2- (2-methoxyethoxy) ethyl ester 110 (13) V-1. Preparation of rhodamine 2- (2-methoxyethoxy) ethyl ester 110 (12) \ 12 To 1.00 grams of rhodamine 110 (2.72 millimoles) was added a mixture of anhydrous DMF and dichloromethane (60 ml: 10 ml), and the mixture was stirred until all the dye was dissolved. 562 milligrams of 1,3-dicyclohexylcarbodi-imide (DCC) (1 equivalent, 2.72 millimoles) were added, followed by 368 milligrams of HOBT (1 equivalent, 2.72 millimoles), 518 microliters of 2- (2-methoxyethoxy) ethanol (1.60 equivalents, 4.36 millimoles), and 33 milligrams (0.27 millimoles) of 4-dimethylaminopyridine (DMAP). The reaction was stirred at room temperature overnight, and then the DMF was distilled off under reduced pressure to give a deep red residue. This residue was subjected to purification by evaporation chromatography using methanol: dichloromethane (2: 8) as eluent, to give (530 milligrams) of a red solid. Thin-layer chromatography (TLC) showed the presence of another product with the desired one. The solid obtained was then dissolved in methanol (10 milliliters), and diethyl ether was added, until a precipitate was obtained. The product was collected and dried to give the desired compound 1_2 (220 milligrams) as a red solid. Rf: 0.33 (MeOH: CH2C12 2: 8). Yield: 18.4 percent. Ms (FAB): Calculated C^ s ^ Os (M-C1) +: 433.1736. Found: 433.1777 V-2 Preparation of 4,5-dibromide-rhodamine 2- (2-methoxyethoxy) ethyl ester 110 (13) 12 13 To a 100 milliliter round bottom flask, equipped with a magnetic stirrer, 235 milligrams (0.50 millimoles) of rhodamine 2- (2-methoxyethoxy) ethylester 110, 122, and 15 milliliters of spectrum-grade methanol were added. The mixture was stirred until all the ro-damine dye was dissolved. Then an amount of 50 microliters (2 equivalents, 1.00 millimoles) of bromine was added, and the reaction was stirred at room temperature for 1 hour and 30 minutes. At the end of the reaction, 10 microliters of cyclohexene was added, and the mixture was stirred for another 10 minutes. The volatile solven-te evaporated under reduced pressure to give a red solid. This solid was passed through chromatography on silica gel using MeOH: CH2C12 (1.2: 8.8) as the eluting solvent. The pure fractions were combined and evaporated to give compound 13 (250 milligrams) as a solid. or Rf: 0.76 (MeOH: CH2C12 2: 8). Yield: 74.3 percent. NMR (CD3OD) d 8.38 (dd, J = 1.5 and 6.87 Hz, 1H); 7.88 (m, 2H); 7. 47 (dd, J = 1.48 and 7.02 Hz, ÍH); 7.15 (d, J = 9.22 Hz, 2H); 7. 04 (d, J = 9.21 Hz, 2H); 4.15 (m, 2H); 3.39-3.25 (, 9H).

Ms (FAB): Calculated for C25H23? 5N2Br2 (M-Br) +: 588.9973. Found: 588.9962 ÜV (MeOH):? Max 502 nm.

EXAMPLE VI VI. Preparation of Rhodamine 3-bromopropylester B (14) To 300 milligrams of rhodamine B (0.62 millimoles), 5 milliliters of dichloromethane was added, and the mixture was stirred until all the dye was dissolved. An amount of 1, 3-dicyclohexylcarbodiimide (DCC) of 142 milligrams (1 equivalent, 0.62 millimoles) was added, followed by 139 milligrams (10.0 millimoles) of 3-bromopropanol, and 8 milligrams (0.06 milligrams). millimoles) of 4-dimethylaminopyridine (DMAP). The reaction was stirred at room temperature overnight. The N, N-dicyclohexylurea was filtered, and the solvent was evaporated in vacuo to give a deep red residue, which was subjected to purification on chromatography by evaporation using methanol: dichloromethane (1: 9) as eluent. The fractions containing the desired compound were combined, and the solvent was evaporated under reduced pressure to give L4 as a deep red viscous solid (300 milligrams). Rf: 0.71 (MeOH: CH2C12 1.5: 8.5). Performance: 79.8 percent. NMR (CD30D) d 8.29 (m, ÍH); 7.85 (m, 2H); 7.43 (m, ÍH); 7.06 (m, 6H); 4.08 (m, 2H); 3.68 (q, J = 7.06 Hz, 8H); 3.21 (m, 2H); 1.81 (m, ÍH); 1.29 (t, J = 7.08 Hz, 12H). Ms (FAB): Calculated C3iH36? 3N2Br2 (M-C1) +: 563.1909. Found: 563.1921 UV (MeOH):? Max 545 nm.

EXAMPLE VII VII. Synthesis of 2,7-dibromo-4 '-carboxytetramethyl-rosamine methyl ester acetate salt (18) VII-1. Preparation of 4'-carboxyhydro-tetramethyl-rosamine methyl ester (17) 910 milligrams of ester 5 (2.08 millimoles) were dissolved in 250 milliliters of dichloromethane and 150 milliliters of water. An excess of NaBH 4 (solid) was added in portions with vigorous stirring, for 30 minutes, until almost all the color was discharged. The pale orange organic phase was separated, and the water phase was extracted twice with dichloromethane. The combined organic layers were dried over Na 2 SO 4, filtered, and evaporated under reduced pressure. The crude oily residue was purified by evaporation chromatography using ethyl acetate as eluent, yielding 530 milligrams of a white foamy solid. Rf: 0.83 (AcOEt). Performance: 63 percent. VII-2. Bromination of dihydro-4 '-carboxytetramethyl-rosamine methyl ester (16) In a 100 milliliter round bottom flask, we introduced 530 milligrams of dihydro-rhodamine hexyl ester B (1.31 ilimoles) and 50 milliliters of spectrum-grade methanol. The mixture was stirred at room temperature until all of the ester dissolved. 2 equivalents of propylene oxide (185 icrolitres, 2.63 millimoles) were added, followed by the dropwise addition of 2 equivalents of bromine (135 microliters, 2.63 millimoles). Stirring was continued at room temperature for 1 hour and 30 minutes. The volatile solvent was evaporated under reduced pressure, and the red oily residue was subjected to purification on chromatography by evaporation using ethyl acetate and hexanes (1: 9) as eluent, to give a white foamy solid (391 milligrams). Rf: 0.36 (AcOEt: Hexanes 1: 9). Yield: 53.5 percent. NMR (CD3OD) d 7.96 (d, J = 8.5 Hz, 2H); 7.28 (d, J = 8.31 Hz, 2H); 7.22 (s, 2H); 6.94 (s, 2H); 3.87 (s, 3H); 2.77 (s, 12H).

VII-3. Oxidation of 2,7-dibromodihydro-4 '-carboxytetramethyl-rosamine methyl ester (17), and formation of the acetate salt of 2,7-dibromo-4'-carboxymethylmethyl-rosamine methyl ester (18) To a stirred solution of 390 milligrams of 2,6-dibromodihydro-tetramethyl-rhodamine methyl ester (0.69 milli-moles) in 15 milliliters of dichloromethane, chlora-nil (1.2 equivalents, 0.83 millimoles, 205 milligrams) was added. The reaction mixture was stirred at room temperature overnight, then the reaction was stopped and the solvent was evaporated under reduced pressure to give a purple residue. The obtained oxidized compound was dissolved in 15 milliliters of dichloromethane, and acetic acid (0.8, milliliters) was added by teo The clear purple solution obtained was stirred for 5 minutes at room temperature, followed by evaporation of the volatile under reduced pressure, to give a purple viscous residue, which was purified by evaporation chromatography using 10 percent methanol in dichloromethane as eluent, to give the desired compound 18A, which is in equilibrium with compound 18B. 18A Rf: 0.34 (MeOH: CH 2 Cl 2 1: 9). 18B Rf: 0.93 (MeOH: CH 2 Cl 2 1: 9). Yield: 30 percent. NMR (CD3OD) d 7.97 (d, J = 8.28 Hz, 2H); 7.45 (d, J = 8.33 Hz, 2H); 7.19 (s, 2H); 6.99 (s, 2H); 3.89 (s, 3H); 2.93 (s, 2.64H); 2.83 (s, 12H); 2.01 (s, 0.356H). Ms (FAB): Calculated for C25H24? 3N2Br2 (MH-AcO) +: 558. 0153. Found: 558 0169 EXAMPLE VIII Preparation of 4,5-dibromo-rhodamine B-lactone (19) ?to 500 milligrams of rhodamine B (1.04 millimoles) were dissolved in 25 milliliters of acetic acid and 25 milliliters of water. Then, 107 microliters of bromine (2 equivalents, 2.08 mmol) was added dropwise, and the reaction mixture was then stirred at room temperature overnight. Water and acetic acid were evaporated under reduced pressure, and the residue obtained was redissolved in dichloromethane and in a 10 percent aqueous solution of sodium bicarbonate. The organic layer was separated and washed twice with water, dried over Na 2 SO 4, filtered and evaporated to give a pink oil. The residue was passed through chromatography on silica gel using methanol: dichloromethane (0.2: 9.8) as eluent, to give 544 milligrams of a white foamy solid. Rf: 0.88 (MeOH: CH 2 Cl 2 1: 9). Performance: 86.8 percent. NMR (CD3OD) d 7.89 (dd, J = 1.45 and 7.8 Hz, ÍH); 7.62 (, 2H); 7. 14 (dd, J = 1.6 and 7.2 Hz, ÍH); 6.81 (d, J = 9.2 Hz, 2H); 6.58 (d, J = 9.2 Hz, 2H); 3.02 (q, J = 7.05 Hz, 8H); 0.93 (t, J = 7.04 Hz, 12H). Ms (FAB): Calculated for C28H2903N2Br2 (MH) +: 599.0545. Found: 599.0527.

EXAMPLE IX Preparation of 2,7-dibromo-rhodamine B-lactone (20) twenty To a stirred solution of the 2,7-dibromodihydro-rhodamine B methyl ester (3), 46 milligrams (0.10 millimoles) in 4 milliliters of dichloromethane, was added chloranil (1.2 equivalents, 0.12 millimoles, 30 milligrams). The reaction mixture was stirred at room temperature overnight, then the reaction was stopped, and the solvent was evaporated under reduced pressure to give a purple residue. The oxidized compound obtained in the previous step was dissolved in 4 milliliters of dioxane, and HCl (IM) (5 milliliters) was added dropwise, and the resulting solution was heated in a water bath to give a clear red solution. After evaporation to dryness under reduced pressure, we obtained a purple viscous residue. The residue was purified by evaporation chromatography using ethyl acetate hexanes (1.5: 8.5) as eluent, to give the desired compound _4 as a white foamy solid (35 milligrams). Rf: 0.34 (AcOEt: hexanes 1.5: 8.5).

Performance: 80 percent. NMR: (CD3OD) d 7.92 (dd, J = 1.45 and 7.5 Hz, ÍH); 7.63 (m, 4H); 7.18 (dd, J = 1.6 and 7.2 Hz, ÍH); 7.02 (m, 2H). Ms (FAB): Calculated for C28H2903N2Br2 (MH) +: 599.0545. Found: 599.0570.

EXAMPLES OF USES OF RODAMINE DERIVATIVES ACCORDING TO THE INVENTION AS INTERMEDIARIES EXAMPLE X X. Synthesis of methyl ester chloride of 4-bromo-5-phenyl-rhodamine B (22) X-1. Preparation of 4-bromo-5-phenyl-rhodamine B-lactone (21) A stirred mixture of 10 millimoles of dibromolacto-na 1 _, 10 millimoles of phenylboronic acid, 4.2 milliliters (30 millimoles) of Et3N, 0.067 grams (0.3 millimoles) of Pd (OAc) 2, and either 0.19 grams (0.62 millimoles) of tri-o-tolylphosphine catalyst, or 0.16 grams (0.62 millimoles) of PPh catalyst, in 40 milliliters of DMF, is under a nitrogen atmosphere at 100 ° C for 2 to 3 hours. The solvent is then distilled under reduced pressure, and the residue is partitioned between CH2C12 and 10 percent aqueous NH3. Then the organic extracts are dried (MgSO 4) and concentrated under reduced pressure. Purification by chromatography by evaporation on silica gel affords the pure monobromolactone 21. (See WJ Thompson and J. Gaudino, J. Org. Chem. 1984, 49, 5237-5243, N. Miyaura, T. Yanagi, and A. Suzuki, Sinthetic Communications, 1981, 11 (7), 513-519).

X-2 Preparation of 4-bromo-5-phenyl-rhodamine B methyl ester chloride (22) 21 ik Methanolysis and concomitant oxidation of monobromolactone 2JL is performed, by first stirring a mixture of 3 to 4 millimoles of the compound in 100 milliliters of methanol, while a fine stream of anhydrous HCl gas is bubbled over a period of 45 minutes, and then heat the mixture to reflux overnight. Then the meta- Nol under reduced pressure, and the dark red residue is purified by evaporation chromatography to provide the desired dark red product 22.

EXAMPLE XI XI. Synthesis of 2,6-dibromo-4,5-dimethyl-rhodamine B (24) XI-1 methyl ester bromide. Preparation of 4,5-dimethyl-rhodamine B-lactone (23) 1S 23 To a solution of 7.0 millimoles of dibromolacto-na 1_9 hexamethylphosphoramide (HMPA), 0.05 millimoles of the benzylbromo-bis (triphenylphosphine) aladin (II) catalyst, and 16.0 millimoles of tetra-methyltin are added. Then the solution is heated to 65 ° C with shaking under air in a sealed tube until the blackening occurs. Then the solution is cooled to room temperature, and 5 milliliters of water are added. The mixture is extracted with dichloromethane, and the organic solution is dried over MgSO4. The evaporation of the solvent produces the crude product, which is purified by evaporation chromatography on silica gel to give the pure lactone 23. (See D. Milstein and J.K. Stille, J. Amer. Chem. Soc. 1979, 101 (17), 4992-4998).

XI-2. Preparation of 2,7-dibromino-4,5-dimetho-rhodamine B methyl ester bromide (24) 2. 3. 4 A solution of 1.25 millimoles of 4,5-dimethyl-rhodamine B-lactone 23 ^ in 50 milliliters of methanol was treated at room temperature with 2.5 millimoles of bromine. A precipitate formed after the addition of the bromine, and the mixture was stirred for 3 hours. The solvent was evaporated under reduced pressure to give a red solid, which was recrystallized from methanol: diethyl ether, to give the desired dibromomethylester 24.

EXAMPLE XII XII. Synthesis of methyl ester bromide of 2-bromo-7-ethynyl-rhodamine B (26) XII-1. Preparation of 2-bromo-7-ethynyl-rhodamine B-lactone (25) 25 A mixture of 53 millimoles of dibromolactone 2_0, and 53 millimoles of ethynyltrimethylsilane, 300 milligrams of triphenylphosphine, and 150 milligrams of palladium (II) acetate, is prepared in 100 milliliters of deaired anhydrous triethylamine at 30 ° C-40 ° C. Then the mixture is heated under argon at 90 ° C-100 ° C for 22 hours. The mixture is cooled and filtered to give the desired impure trimethylsilyl derivative of 25_. Treatment with potassium carbonate at 25 ° C for 16 hours, followed by neutralization, gives lactone 2_5 after purification by evaporation chromatography. (See WB Austin, N. Bilow, WJ Kelleghan, and KSY Lau, J. Org. Chem. 1981, 46, 2280-2286, S. Takahashi, Y. Kuroyama, K. So-nogashira, N. Hagihara, Synthesis, 1980, 627-630).

XII-2. Preparation of 2-bromo-7-ethynyl-rhodamine B methyl ester chloride (26) 2 £ 26 A stirred solution of 3.5 millimoles of 2-bromo-7-ethynyl-rodane B-lactone 2_5 in 100 milliliters of methanol is treated with a fine stream of bubbled HCl gas for 45 minutes. The reaction mixture is then heated to reflux overnight, and the methanol is evaporated under reduced pressure. The dark red residue is purified by evaporation chromatography using a mixture of methanol and dichloromethane to provide the desired ester 26. EXAMPLE XIII XIII. Synthesis of methyl ester bromide of 4,5-dibromo-2,7-di-n-byl-rhodamine B (28) XII-1. Preparation of 2,7-di-n-butyl-rhodamine B-lactone (27) p A solution of 7.0 millimoles of dibromolactone 20 in 4 milliliters of hexamethylphosphoramide (HMPA) is treated with 0.05 millimoles of benzylbromobis (triphenylphosphine) palladium (II), and 16.0 millimoles of a tetra-n-butyltin compound. Then the solution is heated to 65 ° C under air in a sealed tube until the blackening occurs. The solution is then cooled to room temperature, and 5 milliliters of water are added. The mixture is extracted with dichloromethane, and the latter is evaporated in vacuo to give the crude product, which is purified by evaporation chromatography to give the pure lactone 2_7. (See D. Milstein and J. K. Stille, J. ñmer, Chem. Soc. 1979, 101 (17), 4992-4998). XIII-2. Preparation of 4,5-dibromo-2,7-di-n-butyl-rhodamine B methyl ester bromide (28) 21 28 A solution of 1.25 millimoles of rhodamine B-lactone 27 in 50 milliliters of methanol is treated, at room temperature, with 2.5 millimoles of bromine. A precipitate forms shortly after the addition of the bromine, and the mixture is stirred for 3 hours. Then the solvent evaporates under reduced to give a red solid, which is recrystallized from methanol: diethylether, to give the desired di-bromomethylester bromide 28.

DETERMINATION OF BACTERIOSIDIC AND / OR BACTERIOSTATIC RODAMINE DERIVATIVES Experimental Design The following experimental procedures have been used for the determination of antibacterial activity. Bacteriostasis: Escherichia coli: (0157) a The protocol used for bacterial inactivation was performed as described in Brasseur and collaborators, with a few modifications for the evaluation of the bacterioside potential (Brasseur et al., 2000). Compounds TH9402, HA-X-44, HA-X-164, HA-X-171, and HA-VIII-92, showed antibacterial activity against E. coli, using the following experimental procedure. The bacteria were grown overnight in a medium of Lubria broth (LB), 100 microliters (approximately lxlO7 bacteria) of each bacterial suspension was added to 4 milliliters of LB medium, a small aliquot of the bacterial suspension was recovered for the bacilli titration. -terial before treatment, expressed as CFU / ml (united- colony forming per milliliter). The rhodamine derivatives were added to the bacterial suspension, each derivative being tested in duplicate at a concentration of 50 μM. Each mixture of rhodamine-derived bacteria was incubated at 37 ° C for 40 minutes. Then the suspensions of rhodamine bacteria were treated and exposed to light of a wavelength of 514 nanometers for 180 minutes, for a total output energy of 30 Joules / cm2. Following the treatment time, the suspensions of rhodamine bacteria were centrifuged at 3000g, resuspended in 4 milliliters, and serial dilutions were made for each duplicate. Ten microliters of the diluted bacterial suspensions were applied, and the plates were incubated overnight at 37 ° C. The bacteriostatic effect is expressed by the number of colony-forming units per milliliter. Pseudomonas aeruginosa: The protocol used for bactericidal inactivation was performed as described in Brasseur et al., With a few modifications for the evaluation of bacterioside potential (Brasseur et al., 2000). Compound TH9402 showed an antibacterial activity against P. aeruginosa using the following experimental procedure. The bacteria were grown overnight in a medium of Lubria broth (LB), 100 microliters were added (approximately lxlO7 bacteria) from each bacterial suspension to 4 milliliters of LB medium, a small aliquot of the bacterial suspension was recovered for bacterial titration before treatment, expressed as CFU / ml. The rodamine derivatives were added to the bacterial suspension, each derivative being tested in duplicate at a concentration of 50 μM. Each mixture of bacteria-derivative of roda-mina was incubated at 37 ° C for 40 minutes. Then the suspensions of rhodamine bacteria were treated and exposed to light of a wavelength of 514 nanometers for 180 minutes, for a total output energy of 30 Joules / cm2. Following the treatment time, the suspensions of rhodamine bacteria were centrifuged at 3000g, resuspended in 4 milliliters, and serial dilutions were made for each duplicate. Ten microliters of the diluted bacterial suspensions were applied, and the plates were incubated overnight at 37 ° C. The bacteriostatic effect is expressed by the number of colony forming units per milliliter.

Salmonella typhlmurxtim. The protocol used for bacterial inactivation was carried out as described in Brasseur et al., With a few modifications for the evaluation of the bacterioside potential (Brasseur et al., 2000).

The compounds PH9402, HA-X-44 and HA-X-164 showed antibacterial activity against S. typhimurium using the following experimental procedure. The bacteria were grown overnight in a medium of Lubria broth (LB), 100 microliters (approximately lxlO7 bacteria) from each bacterial suspension was added to 4 milliliters of LB medium, a small aliquot of the bacterial suspension was recovered for bacterial titration before treatment, expressed as CFU / ml. The rodamine derivatives were added to the bacterial suspension, each derivative being tested in duplicate at a concentration of 50 μM. Each mixture of bacteria-derivative of roda-mina was incubated at 37 ° C for 40 minutes. Then the suspensions of rhodamine bacteria were treated and exposed to light of a wavelength of 514 nanometers for 180 minutes, for a total output energy of 30 Joules / cm2. Following the treatment time, the suspensions of rhodamine bacteria were centrifuged at 3000g, resuspended in 4 milliliters, and serial dilutions were made for each duplicate. Ten microliters of the diluted bacterial suspensions were applied, and the plates were incubated overnight at 37 ° C. The bacteriostatic effect is expressed by the number of colony forming units per milliliter.

Staphylococcus epxdermitis The protocol used for bactericidal inactivation was performed as described in Brasseur et al., With a few modifications for the evaluation of bacterioside potential (Brasseur et al., 2000). Compounds TH9402, HA-X-40, HA-X-44, HA-X-149, and HA-X-164 showed antibacterial activity against S. epidermitis using the following experimental procedure. Bacteria were grown overnight in a medium of Lubria broth (LB), 100 microliters (approximately IxlO7 bacteria) from each bacterial suspension was added to 4 milliliters of LB medium, a small aliquot of the bacterial suspension was recovered for bacterial titration before treatment, expressed as CFU / ml. The rodamine derivatives were added to the bacterial suspension, each derivative being tested in duplicate at a concentration of 50 μM. Each mixture of bacteria-derivative of roda-mina was incubated at 37 ° C for 40 minutes. Then the suspensions of rhodamine bacteria were treated and exposed to light of a wavelength of 514 nanometers for 180 minutes, for a total output energy of 30 Joules / cm2. Following the treatment time, the suspensions of rhodamine bacteria were centrifuged at 3000g, resuspended in 4 milliliters, and serial dilutions were made for each duplicate. 10 microliters of the sus- Diluted bacterial pensions, and the plates were incubated overnight at 37 ° C. The bacteriostatic effect is expressed by the number of colony forming units per milliliter. Staphylococcus epidexmitis The protocol used for bactericidal inactivation was carried out as described in Brasseur et al., With a few modifications for the evaluation of the bacterioside potential (Brasseur et al., 2000). Compounds HA-X-171 and HA-VIII-92 showed antibacterial activity against S. epidermitis using the same experimental procedure, except that a concentration of 10 μM was used in the experimental procedure. The bacteria were grown overnight in a medium of Lubria broth (LB), 100 microliters (approximately lxlO7 bacteria) from each bacterial suspension was added to 4 milliliters of LB medium, a small aliquot of the bacterial suspension was recovered for bacterial titration before treatment, expressed as CFU / ml. The rodamine derivatives were added to the bacterial suspension, each derivative being tested in duplicate at a concentration of 50 μM. Each mixture of bacteria-derivative of roda-mina was incubated at 37 ° C for 40 minutes. Then the suspensions of rhodamine bacteria were treated and exposed to a light of a wavelength of 514 nanometers for 180 mi- ñutos, for a total output energy of 30 Joules / cm2. Following the treatment time, the suspensions of rhodamine bacteria were centrifuged at 3000g, resuspended in 4 milliliters, and serial dilutions were made for each duplicate. Ten microliters of the diluted bacterial suspensions were applied, and the plates were incubated overnight at 37 ° C. The bacteriostatic effect is expressed by the number of colony forming units per milliliter. Staphylococcus epidermitls The protocol used for bactericidal inactivation was carried out as described in Brasseur et al., With a few modifications for the evaluation of the bacterioside potential (Brasseur et al., 2000). The compound HA-X-40 showed antibacterial activity against S. epidermitis using the same experimental procedure, except that an extrusion time of 90 minutes was performed before the radiation treatment. The . epidermitis was grown overnight in a medium of Lubria broth (LB), and 100 microliters (lxlO7 bacteria) of each bacterial suspension was added to 4 milliliters of LB medium. A small aliquot of the bacterial suspension was recovered for bacterial titration before treatment. The rhodamine derivatives were added to the bacterial suspension, the derivative being tested in duplicate. in a concentration of 50 μM. Each mixture of rhodamine-derived bacteria was incubated at 37 ° C for 40 minutes. The bacterial suspensions were then centrifuged at 3000g for 10 minutes, and resuspended in 4 milliliters of the LB medium, and incubated for 90 minutes at 37 ° C to allow the extrusion of the derivatives. Then the suspensions of bacteria-rhodamines were treated and exposed to light of a wavelength of 514 nanometers for 180 minutes, for a total output energy of 30 Jou-les / cm2. Following the treatment time, serial dilutions were made for each duplicate, and 10 microliters of the diluted bacterial suspension was applied, and the plates were incubated overnight at 37 ° C. The bacteriostatic effect is expressed by the number of colony forming units / milliliter.

DETERMINATION OF THE ANTIVIRAL ACTIVITY OF THE DERIVATIVES OF RODAMINE OF FORMULA (I) Antiviral Assay: References: Lin L., Inactivation of cytomegalovirus in platelet concentrates using Helinx ™ technology, Seminar in Hematology, 2001, 38, 4, Sup. # 11, 27-33; Brasseur, N., Ménard, I., Forget, A., El Jastimi, R., Hamel, R., Molfino, N.A. and E van Lier, J., Eradication of Multiple Myeloma and Breast Cancer Cells by TH9402-mediated Photodynamic Therapy: Implication of Clinical Ex vivo Purging of Autologous Stern Cell Transplant, Photochemistry and Pho-tobiology, 2000, 72, 6, 780-878; Lin. B., Londe, H., Janda, J.M. , Hanson, C.V. and Corash, L., Photochemical Inactivation of Pathogenic, Bacteria in Human Platelet Concentrates, Blood, 1994, 83, 9, 2698-2706; Lin. BL, Londe, H., Hanson, CV, Wiesehahn, G., Isaacs, S., Cimino, G. and Corash, L., Photochemical Inactivation of Cell-Associated Human Immunodeficiency Virus in Platelets Concentrates, Blood, 1993, 82, 1, 292-297; Lin, B.L., Wiesehahn, G.P., Morel, P.A. and Corash, L., Use of 8-Methoxypsoralen and Long-Wavelegth Ultraviolet for Decontamination of Platelet Concentrates, Blood, 1989, 74, 1, 517-525. Objective: The antiviral assay was performed as described in Lin, L (2001). Human diploid fibroblasts, prepuce cells (FS), were used in this assay. The antiviral activity of the rhodamine derivatives was tested, and the results showed that all the compounds, HA-X-40, HA-X-149, HA-X-164, HA-X-171, and HA-VIII -92, followed by PDT treatment, possess antiviral activity against cytomegalovirus. Method: Prepuce cells were cultured until the creep in covered jars. At the time of infection, 2.5-3.5xl05 cells were being cultured on each slide. The CMV supply solution (AD169) containing 1 milliliter of the virus, thawed rapidly, braked, and diluted following 100-fold dilutions in MEM (Earle's salt) supplemented with L-glutamine and 2 percent calf fetal serum, in a total volume of 30 milliliters. The titration of the virus has been determined in 10-2 (104 TCID50) plaque forming units (pfu) in 0.2 milliliters. Therefore, 1 milliliter used in the PDT experiments represents 1.4xl05 plaque forming units. A.M. O. I. from 0.4 to 0.5 of CMV was used throughout this experiment. The plates that did not contain rhodamine derivatives were treated with light in parallel with the plates not treated with light. The concentration used throughout the assay for rhodamine derivatives was maintained at 50 μM. Following the addition of the derivatives to the viral delivery solution, the plates were placed in the The-ralux L6.30 device and illuminated for 180 minutes with 200 rpm shaking. The energy output was measured at 30 Joules / cm2. The plate without PDT was placed in an incubator at 37 ° C for the same amount of time. Following this time of treatment, dilutions were made and inoculated with the foreskin cells under centrifugation (2000g, 60 minutes). Following the centrifugation, the cells were incubated for 60 minutes at 37 ° C, with 5% CO2, then washed with the culture medium, and incubated for 18 to 24 hours at 37 ° C with 5% CO 2. percent. The volume of inoculation of each dilution was 0.2 milliliters, and the dilutions made were from 10 ~ 3 to 10_s in duplicate. The cells were fixed, removed from the flasks, and stained with the immediate early antigen of the monoclonal antibody to cytomegalovirus with labeled FITC (fluorescein isothiocyanate). Cytomegalovirus viral particles were counted. A fluorescent virus particle (in the form of a kidney) represents a plaque forming unit. Here is the protocol for the other two compounds TH9402 and HA-X-44, which inhibit cytomegalovirus infectivity without treatment with PDT, as well as a new version of the table, to be added in the patent. Objective: The antiviral assay was performed as described in Lin, L. (2001). Human diploid fibroblasts, prepuce cells (FS), were used in this assay. The antiviral activity of the rhodamines was tested, and the results showed that the compounds TH9402 and HA-X-44 did not need treatment with PDT to possess antiviral activity against the cytometry. galovirus. Method: Foreskin cells were grown to confluence in covered flasks. At the time of infection, 2.5-3.5xl05 cells were being cultured on each slide. The CMV supply solution (AD169) containing 1 milliliter of the virus was thawed rapidly, seeded, and diluted following 100-fold dilutions in MEM (Earle's salt) supplemented with L-glutamine and fetal calf serum at 2 times cent, in a total volume of 30 milliliters. The titration of the virus has been determined in 10"2 (104 TCID50) plaque forming units (pfu) in 0.2 milliliters, therefore 1 milliliter used in the PDT experiments represents 1.4x10 plaque forming units. 0.5 of CMV was used throughout this experiment.The plates that did not contain rhodamine derivatives were treated with light in parallel with the plates not treated with light.The concentration used throughout the trial for rhodamine derivatives was maintained in 50 μM Following the addition of the derivatives to the viral delivery solution, the dilutions were made and inoculated with the foreskin cells under centrifugation (2000g, 60 minutes). incubated They were rinsed for 60 minutes at 37 ° C, with C02 at 5 percent, then washed with the culture medium, and incubated for 18 to 24 hours at 37 ° C with 5 percent C02. The volume of inoculation of each dilution was 0.2 milliliters, and the dilutions made were 10 ~ 3 to 10"5 in duplicate.The cells were fixed, removed from the flasks, and stained with the immediate early antigen of the monoclonal antibody to cytomegalovirus with labeled FITC (fluorescein isothiocyanate) Cytomegalovirus viral particles were counted One particle of fluorescent virus (kidney-shaped) represents a plaque-forming unit Although the invention has been described in connection with the modalities specific thereto, it will be understood that further modifications may be made, and it is intended that this application cover any variations, uses, or adaptations of the invention by generally following the principles of the invention, and including departures from the present disclosure entering within the known or customary practice in the art to which the invention pertains, and how it can be applied to the the essential features stipulated hereinbefore, and as follows in the scope of the appended claims.

Claims (50)

1. CLAIMS Rhodamine derivatives of the formula (I) ? wherein: - one of Ri, R2, R3, R, and Rio represents a halogen atom, and each of the remaining Ri, R2, R3, R, and each of the remaining Rio group, is independently selected from group consisting of hydrogen, halogen atoms, an amino group, acylamino, dialkylamino, cycloalkylamino, azacycloalkyl, alkylcycloalkylamino, aroylamino, diarylamino, arylalkylamino, aralkylamino, alkylarylamino, arylaralkylamino, hydroxy, alkoxy, aryloxy, aralkyloxy, mercapto, thioalkyl , thioaryl, thioaralkyl, carboxyl, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, no, hydroxysulfonyl, amidosulfonyl, dialkylamidosulfonyl, arylalkylamidosulfonyl, formyl, acyl, aroyl, alkyl, alkylene, alkenyl, aryl, aralkyl, vinyl, alkynyl, and the corresponding substituted groups; - m = 0-1; n = 1-4; A is nothing, O, or NH; Rg represents an alkylene group; Z is H, amino, dialkylamino, or a trialkylamino salt; - X "is an anion, and Rs, Re, R, and R8 are independently H or alkyl of 1 to 6 carbon atoms, or Ri in combination with R5 or Re, or R2 in combination with 5 or Re, or R3 in combination with R7 or R8, or R in combination with R or R8, represents an alkylene group, alone or in association with a pharmaceutically acceptable carrier, with the proviso that the following specific compounds are excluded: 4,5-dibromo-rhodamine 123 '(2- (4,5-dibromo-6-amino-3-imino-3H-xanten-9-yl) -benzoic acid methyl ester hydrochloride) - 4,5-dibromo-rhodamine 123 (ethyl hydrochloride) 2- (4,5-dibromo-6-amino-3-imino-3H-xanten-9-) ester il) -benzoic acid); 4, 5-dibromo-rhodamine 123 (2- (4,5-dibromo-6-amino-3-imino-3 H-xanten-9-yl) -benzoic acid octylester hydrochloride); - 4,5-dibromo-rhodamine 110-n-butylester 110 (2- (4,5-dibromo-6-amino-3-imino-3 H-xanten-9-yl) -benzoic acid n-butylester hydrochloride); and rhodamine n-butylester B (2- (6-ethylamino-3-ethylimino-3H-xanten-9-yl) -benzoic acid n-butylester hydrochloride).
2. 2. Derivatives of rhodamine according to claim 1, wherein: "alkyl" means a straight or branched chain aliphatic hydrocarbon group, and the corresponding substituted alkyl group bearing one or more substituents which may be the same or different, and which are selected from the group consisting of halogen, aryl, hydroxyl, alkoxy, aryloxy, alkyloxy, thioalkyl, thioaryl, aralkyloxy, thioaralkyl, and cycloalkyl, and "branched" means a lower alkyl group, such as methyl, ethyl, or propyl, is attached to a linear alkyl chain, and preferred alkyl groups include the "lower alkyl" groups, which are the alkyl groups having from about 1 to about 6 carbon atoms, and the alkyl groups of example are the methyl, ethyl, isopropyl groups, hexyl, cyclohexylmethyl, and methyl or ethyl groups are more preferred; "cycloalkyl" means a non-aromatic ring preferably composed of 4 to 10 carbon atoms, and the cyclic alkyl group may be partially unsaturated, and preferred cyclic alkyl rings include cyclopentyl, cyclohexyl, cyclo-octyl; the cycloalkyl group may be optionally substituted with an aryl group substituent, and the cyclopentyl and cyclohexyl groups are preferred; "alkenyl" means an alkyl group containing a carbon-carbon double bond, and preferably having from 2 to 5 carbon atoms in the straight chain, and the example groups include allylvinyl; "alkynyl" means an alkyl group containing a carbon-carbon triple bond, and preferably having from 2 to 5 carbon atoms in the straight chain, and the example groups include ethynyl and propargyl; "aryl" means an aromatic carbocyclic radical or a substituted carbocyclic radical preferably containing from 6 to 10 carbon atoms, such as phenyl or naphthyl, or phenyl or naphthyl substituted by at least one of the substituents selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, aralkyl, hydroxyl, alkoxy, aryloxy, aralkoxy, carboxyl, aroyl, halogen, nitro, trihalomethyl, cyano, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acylamino, aroylamino, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, thioalkyl, thioaryl, alkylene, or -NYY ', wherein Y and Y' are independently hydrogen, alkyl, aryl, or aralkyl; "aralkyl" means a radical in which an aryl group is substituted by an alkyl H atom, and the example aralkyl group is benzyl; "acyl" means an alkyl-CO- group, wherein the alkyl group is as described above, and the preferred acyl has an alkyl group containing from 1 to 3 carbon atoms in the alkyl group, and the example groups include acetyl, propanoyl, 2-methylpropanoyl, bu-tanoyl, or palmitoyl; "Aroyl" means an aryl-CO- group, in which the aryl group is as described above, and preferably contains from 6 to 10 carbon atoms in the ring, and the example groups include benzoyl and 1- and 2-naphthoyl; "alkoxy" means an alkyl-O- group, wherein the alkyl group is as described above, and the example alkoxy groups include methoxy, ethoxy, normal propoxy, isopropoxy, normal butoxy, and heptoxy; "aryloxy" means an aryl-O- group, wherein the aryl group is as described above, and the exemplary aryloxy groups include phenoxy and naphthoxy- the; "thioalkyl" means an alkyl-S- group, wherein the alkyl group is as described above, and the exemplary thioalkyl groups include thiomethyl, thioetyl, thioisopropyl, and thioheptyl; "thioaryl" means an aryl-S- group, wherein the aryl group is as described above, and the exemplary thioaryl groups include thiophenyl and thionaphthyl; "aralkyloxy" means an aralkyl-0- group, wherein the aralkyl group is as described above, and the example aralkyloxy group is benzyloxy; "thioaralkyl" means an aralkyl-S- group, wherein the aralkyl group is as described above, and the example thioaralkyl group is thiobenzyl; "dialkylamino" means a group -NYY ', wherein both Y and Y' are alkyl groups as described above, and example alkylamino groups include ethylamino, dimethylamino, and diethylamino; "alkoxycarbonyl" means an alkyl-0-CO- group, wherein the alkyl group is as described above, and the example alkoxycarbonyl groups include methoxy- and ethoxycarbonyl; - "aryloxycarbonyl" means an aryl-0- group CO-, wherein the aryl group is as described above, and exemplary aryloxycarbonyl groups include phenoxy- and naphthoxycarbonyl; "aralkoxycarbonyl" means an aralkyl-0-CO- group, wherein aralkyl is as defined above, and the exemplified aralkoxycarbonyl group is benzyloxycarbonyl; "carbamoyl" is a group H2N-CO-; "alkylcarbamoyl" is a group Y'YN-CO-, wherein one of Y and Y 'is hydrogen, and the other of Y and Y' is alkyl as defined above; "dialkylcarbamoyl" is a group Y'YN-CO-, wherein both Y and Y 'are alkyl as defined above; - "acylamino" is an acyl-NH group, wherein acyl is as defined above; "aroilamino" is an aroyl-NH group, where aroyl is as defined above; "alkylene" means a straight or branched chain bivalent hydrocarbon group preferably having from 2 to 8 carbon atoms, and the alkylene group may be interrupted by one or more substituted nitrogen atoms, wherein the substituent of the nitrogen atom is alkyl or oxygen or sulfur atoms, and it is currently preferred more than the alkylene group to have 2 to 3 carbon atoms. carbon, and the example alkylene groups include ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), -CH2NMe-CH2-, 0-CH2-0, or -0-CH2CH2-0-; "halogen" preferably means fluorine, chlorine, bromine, or iodine; "Azacycloalkyl" preferably means a saturated carbon ring of 4 to 9 members, wherein one of the methylene groups is replaced by nitrogen; "cycloalkylamine" means a group -NYY ', wherein one of Y and Y is hydrogen, and the other of Y and Y is cycloalkyl as defined above; "alkylcycloalkylamino" means a group -NYY ', wherein one of Y and Y' is alkyl as defined above, and the other of Y and Y 'is cycloalkyl as defined above; "diarylamino" means a group -NYY ', wherein both Y and Y' are aryl groups as described above; "aralkylamino" means a group -NYY ', wherein one of Y and Y' is hydrogen, and the other of Y and Y 'is aralkyl as defined above; "arylalkylamino" means a group -NYY ', wherein one of Y and Y' is alkyl as defined above, and the other of Y and Y is aryl as defined above; - "alkylalkylamino" means a group - NYY ', wherein one of Y and Y' is alkyl as defined above, and the other of Y and Y 'is aralkyl as defined above; "arylalkylamino" means a group -NYY ', wherein one of Y and Y' is aryl as defined above, and the other of Y and Y 'is aralkyl as defined above; "mercapto" is a group -SH or SR, wherein R can be any of the Ri to Rio groups previously de-finidos, -SH, mercaptoaryl, and mercaptoalkyl groups are preferred; "hydroxysulfonyl" is -S03H; "amidosulfonyl" is -S02NH2; "dialkylamidosulfonyl" means a group -S02NYY ', wherein both Y and Y' are alkyl groups as described above; "arylalkylamidosulfonyl" means a group -S02NYY ', wherein one of Y and Y' is aryl as defined above, and the other of Y and Y 'is aralkyl as defined above; and - "anion" means the deprotonated form of an organic or inorganic acid, and the anion is preferably selected from hydrochlorides, hydrobromides, sulfates, nitrates, borates, phosphates, oxalates, tartrates, maleates, citrates, acetates, ascorbates, succinates, benzenesulfonates, methanesulfonates, cyclohexansulfonates, toluenesulfonates, sulfamates, lactates, malonates, ethanesulfonates, cyclohexyl sulphonates, and kinetics. In the case where the rodamine derivative carries one or more acid substituents, the compound covered includes the inner salt or any salt derived from neutralization by any of the following bases: sodium hydroxide, potassium hydroxide, calcium hydroxide, lithium hydroxide, ammonia, ethylenediamine, lysine, diethanolamine, piperazine, and the like.
3. 3. Derivatives of rhodamine according to claim 1 6 2, wherein at least two of the Ri / R2 groups. R3. R. and Rio represent a halogen atom, which preferably is a bromide atom.
4. 4. Rhodamine derivatives according to any of claims 1 to 3, wherein the halogen atoms are in the 2-7, 4-5, or 4 '-5' position, on the ring, or are the end of the ester chain.
5. 5. Methyl ester hydrochloride of 2'- (6-dimethylamino-3-dimethylimino-3H-xanten-9-yl) - ', 5'-dichloro-benzoic acid. 6. 2- (2-methoxyethoxy) ethyl ester of 4,5-dibromo-rhodamine 110. 7. Hexyl ester acetate salt of 2,7-dibromo-rhodamine B. 8. 2,7-dibromo methyl ester acetate salt - rhodamine B 9. 4, 5-dibromo-rhodamine 6G. 10. Rhodamine 3-bromopropylester B. 11. Intermediates represented by one of formulas (II) to (VII) and (I '), as defined in the Figures 1 to 5, wherein the different groups are as defined any of claims 1 to 4. 12. Processes for the synthesis of new rhodamine derivatives of the formula (I), wherein the different R a Ro, A, X, Y, Y 'and Z, myn, are defined as in claims 1 to 10, without the exclusion of the compounds listed in the condition at the end of claim 1, as defined in the schemes depicted in Figures 1 to 5, and in the corresponding parts of the description. 13. The use of the rhodamine derivatives of the formula (I), as defined in any of claims 1 to 10, in the treatment of immune disorders. 14. Photoactivatable rhodamine derivatives to improve a high yield of quantum yield, and the generation of singlet oxygen, after irradiation, while maintaining a desirable differential retention of rhodamine between normal and cancer cells, being these derivatives as defined in any of claims 1 to 10. 15. The use of photoactivatable derivatives as defined in any of claims 1 to 10, without the exclusion of the compounds listed in the condition at the end of claim 1, for the photodynamic therapy of cancer patients, by the destruction of human cancer cells, where they are achieved appropriate intracellular levels of these derivatives, and irradiation of an appropriate wavelength and intensity is applied. 16. A method for photodynamic therapy of patients suffering from leukaemias, multiple disseminated myelomas, or lymphomas, which comprises the steps of: a) harvesting the patient's human bone marrow; b) purging the bone marrow from step a) by photodynamic therapy, using a therapeutic amount of a photoactivatable derivative according to any one of claims 1 to 10, without excluding the compounds listed in the condition at the end of claim 1, under irradiation of a suitable wavelength; and c) performing autologous stem cell transplantation using the purged bone marrow from step b). The method of claim 16, wherein the purging of step b) further comprises intense chemotherapy and total body irradiation (TBI) procedures. 18. A method for in vitro purging of human bone marrow containing solid tumor metastases, selected from the group consisting of metastases from chest, lung, prostate, pancreatic, and of Edenic carcinomas, melanomas and disseminated sarcomas, where surgical separation and volume reduction can be achieved, which comprises the steps of: a) harvesting the patient's human bone marrow; b) purging the bone marrow from step a) by photodynamic therapy, using a therapeutic amount of at least one photoactivatable derivative according to any of claims 1 to 10, without excluding the compounds listed in the condition at the end of the claim 1, under irradiation of a suitable wavelength; and c) performing autologous stem cell transplantation using the purged bone marrow from step b). The method of claim 18, wherein the purging of step b) further comprises intense chemotherapy and total body irradiation (TBI) procedures. 20. A method for photodynamic therapy of cancer patients, which comprises administering to these patients a therapeutically acceptable intracellular level of at least one photoactivatable derivative according to claims 1 to 10, without the exclusion of the compounds listed in FIG. the condition at the end of claim 1, and subjecting these patients to irradiation of a therapeutically adequate wavelength. 21. The method of claim 20, wherein the photoactivatable derivative is administered by instillation, injection, diffusion into the bloodstream at tumor sites directly accessible to light emission, or at tumor sites accessible to the laser beams, using rigid or flexible endoscopes. 22. The method of claim 21, wherein the laser-accessible tumor site is selected from the group consisting of urinary bladder, oral cavity, esophagus, stomach, lower digestive tract, upper and lower respiratory tract. 23. A method for the treatment of leukemias in patients, which comprises the steps of: a) purging cancerous clones from the patients' bone marrow; b) subjecting the purged clones of step a) to a photodynamic treatment using a therapeutic amount of at least one of the photoactivatable derivatives according to claims 1 to 10, without excluding the compounds listed in the condition at the end of the claim 1, under irradiation of a suitable wavelength, for the selective destruction of the leukemic cells, without affecting the normal cells of the patients; and c) administering the treated clones of step b) to patients; without causing in this way a systemic toxicity for patients. 24. A photoactivatable pharmaceutical composition for the selective destruction and / or inactivation of immunologically reactive cells, unaffected to normal cells, and without causing systemic toxicity to the patient, this composition comprising at least one photoactivatable whey derivative or a salt thereof, as defined in claims 1 to 10, without the exclusion of the compounds listed in the condition at the end of claim 1, and photoactivatable derivatives thereof; in association with a pharmaceutically acceptable vehicle; wherein the photoactivation of these derivatives induces the annihilation of the cells, while the inactivated derivatives are substantially non-toxic to the cells. 25. The use of the photoactivatable derivatives of claim 1, for the photodynamic treatment for the selective destruction and / or inactivation of the immunologically reactive cells, without affecting the normal cells, and without causing systemic toxicity for the patient, in where appropriate intracellular levels of these derivatives are reached, and irradiation of an appropriate wavelength and intensity is applied. 26. A method of preventing graft-versus-host disease associated with cell transplantation those of allogeneic stem in a patient, which comprises the steps of: a) activating the lymphocytes of a donor, mixing the cells of the donor with the cells of the host for a sufficient time for a sufficient period of time to be present an immune reaction; b) substantially removing activated lymphocytes from step a) with photodynamic therapy, using a therapeutic amount of a photoactivatable composition of claim 24, under irradiation of a suitable wavelength; and c) carrying out transplantation of allogeneic stem cells using the treated mixture from step b). 27. A method for the treatment of an immune disorder in a patient, which comprises the steps of: a) harvesting the patient's hematopoietic cells; b) treating the hematopoietic cells ex vivo from step a) by photodynamic therapy, using a therapeutic quantity of a photoactivatable composition of claim 24, under irradiation of a suitable wavelength; and c) performing the graft infusion or autograft transplant, using the treated hematopoietic cells of step b). 28. The method of claim 27, wherein the immunological disorder is selected from the group consisting of conditions wherein the self cells or donor cells react against host tissues or foreign targets, such as graft disease against the host. host, graft rejection, autoimmune disorders, and T-cell mediated immunoallergies. 29. The method of claim 27, wherein the hematopoietic cells are selected from the group consisting of bone marrow, peripheral blood cells and cells. spinal cord blood mononuclear cells. 30. The use of at least one rhodamine derivative or a salt thereof, as defined in any of claims 1 to 10, without the exclusion of the listed compounds in the condition at the end of claim 1, alone or in combination with a vehicle, for the treatment of infections generated by Gra ty / or Gram- bacteria. 31. The use according to claim 30, for the treatment of infections generated by Staphylococcus cider epidermitis. 32. The use of 4,5-dibromo-rhodamine 110 2- (2-methoxyethoxy) ethyl ester, as a bacteriostatic agent against Escherichia coli 0157: H7, and / or against Salmonella typhimurium LT2. 33. The use of the hexyl ester acetate salt of 2, 7-dibromo-rhodamine B, as a bacteriostatic agent against Salmonella typhimurium LT2. 34. The use of 4, 5-dibromo-rhodamine 6G, as a bacteriostatic agent against Escherichia coli 0157: H7. 35. The use of 3-bromopropylester of rhodamine B, as a bacteriostatic agent against Escherichia coli 0157: H7. 36. The use of 4,5-dibromo-rhodamine methyl ester as a bacteriostatic agent against Escherichia coli 0157: H7, Salmonella typhimurium LT2, and / or Pseudomonas aeruginosa. 37. The use according to any of claims 30 to 36, wherein the vehicle is a pharmaceutically acceptable carrier, and is preferably selected from the group consisting of 5 percent mannitol in water, and / or dimethyl sulfoxide. , or any appropriate excipient. 38. A bactericidal composition for the treatment of a liquid (such as a cell suspension, derived from blood, or any other liquid with or without cells) contaminated with Gram + and / or Gram- bacteria, which composition comprises an effective amount of at least a derivative of rhodamine or a salt thereof, as defined in any of claims 1 to 10, without the exclusion of the compounds listed in the condition at the end of the claim 1, alone or in combination with a vehicle. 39. A bactericidal solution, for the treatment of a site contaminated with Gram + and / or Gram- bacteria, whose solution comprises an effective amount of at least one rhodamine derivative, or a salt thereof, as defined in any of the claims 1 to 10, without any knowledge, alone or in combination with a vehicle, for the treatment of infections generated by Gram + and / or Gram- bacteria. 40. A method for the treatment of infections generated by Gram + and / or Gram- bacteria, which method comprises administering to a human or animal that needs it, an effective amount of at least one rhodamine derivative or a salt thereof, such as is defined in any of claims 1 to 10, without any knowledge, alone or in combination with a vehicle. 41. A method according to claim 40, wherein the effective amount administered is between 0.5 and 200 milligrams per kilogram of body weight per day. 42. A medicament containing an effective amount of at least one rhodamine derivative or a salt thereof, as defined in any one of claims 1 to 10, without the exclusion of the compounds listed in the condition at the end of claim 1 , alone or in combination with a vehicle, for the treatment of infections generated by Gram + and / or Gram- bacteria. 43. The use of an effective amount of at least one rhodamine derivative or a salt thereof, as de-fine in any of claims 1 to 10, without any knowledge, alone or in combination with a vehicle, in the treatment of diseases generated by enveloped viruses or by non-enveloped viruses. 44. The use according to claim 43, wherein the enveloped virus is one with a double-stranded DNA, preferably one of the Herpes viridae family. 45. The use of 4,5-dibromo-rhodamine-2 '- (6-dimethylamino-3-dimethylimino-3H-xanten-9-yl) -4', 5'-dichloro-benzoic acid methyl ester hydrochloride, as an antiviral agent against cytomegalovirus. 46. The use of 2,7-dibromo-rhodamine B hexyl ester acetate salt, as an antiviral agent against cytomegalovirus. 47. Hexyl ester acetate salt of 2,7-dibromo-rhodamine B, as an antiviral agent against cytomegalovirus. 48. The 2- (2-methoxyethoxy) ethyl ester of 4,5-dibromo-rhodamine 110, as an antiviral agent against cytomegalovirus. 49. The 4,5-dibromo-rhodamine methyl ester, as a antiviral agent against cytomegalovirus. 50. A medicament containing an effective amount of at least one rhodamine derivative or a same salt, as defined in any of claims 1 to 10, without the exclusion of the compounds listed in the condition at the end of claim 1, alone or in combination with a vehicle, for the treatment of viral infections. SUMMARY Novel compounds of the formula (I) wherein: one of R1, R2, R3, R4, and (R10) n represents a halogen atom, and each of the remaining R1, R2, R3, R4, and each one the remaining RIO group, is independently selected from the group consisting of hydrogen, halogen atoms, an amino group, acylamino, dialkylamino, cycloalkylamino, azacycloalkyl, alkylcycloalkylamino, aroylamino, diarylamino, arylalkylamino, aralkylamino, alkylarylakylane , arylaralkylamino, hydroxy, alkoxy, aryloxy, aralkyloxy, mercapto, thioalkyl, thioaryl, thioaralkyl, carboxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, cyano, hi-droxysulfonyl, amidosulfonyl, dialkylamidosulfonyl, arylalkyl amidosulfonyl, formyl, acyl , aroyl, alkyl, alkylene, alkenyl, aryl, aralkyl, vinyl, alkynyl, and the corresponding substituted groups; - m = 0-1; n = 1-4, A is nothing, O, or NH; R9 represents an alkylene group; -Z is H, amino, dialkylamino, or trialkylamino salt; X- is an anion; R5, R6, R7, and R8 are independently H or alkyl of 1 to 6 carbon atoms, or R1 in combination with R5 or R6, or R2 in combination with R5 or R6, or R3 is combination with R7 or R8, or R4 in combination with R7 or R8, represents an al-kinylene group, alone or in association with a pharmaceutical carrier- mind acceptable. These compounds are useful as intermediates and as bactericides, as well as antiviral agents and in the treatment of immune disorders. General synthesis of substituted rhodamine 4- and 2,7-halogenated derivatives. (I)