AU2002332608A1

AU2002332608A1 - Porphyrins with enhanced multi-photon absorption cross-sections for photodynamic therapy

Info

Publication number: AU2002332608A1
Application number: AU2002332608A
Authority: AU
Inventors: Eric Nickel; Aleksander Rebane; Charles W. Spangler
Original assignee: Montana State University
Current assignee: Montana State University
Priority date: 2001-08-22
Filing date: 2002-08-22
Publication date: 2003-06-05
Anticipated expiration: 2022-08-22

Description

PORPHYRINS WITH ENHANCED MULTI-PHOTON ABSORPTION CROSS-SECTIONS FOR PHOTODYNAMIC THERAPY

This application claims priority under 35 U.S.C. § 119 to U.S. Provisional Patent Application No. 60/313,815 filed on August 22, 2001, and to U.S. Provisional Patent Application No. 60/348,393 filed on January 16, 2002, the disclosures of which are hereby incorporated by reference in their entireties.

BACKGROUND OF THE INVENTION Field ofthe Invention

This invention relates to porphyrin-based compounds modified with a chromophore useful in photodynamic therapy ("PDT"), particularly multi- photon PDT, and methods of using the compounds in PDT and related applications. In particular, a method of treating a tumor disease or a tumor by contacting an therapeutically effective amount of the porphyrin-based compound and irradiating the modified porphyrin with sufficient radiation to produce singlet oxygen is disclosed. Preferably the modified porphyrin simultaneously absorbs two photons of radiation which radiation is preferably is in a range of about 700 nm to about 1300 nm.

Discussion ofthe Related Art

Photodynamic therapy is an accepted treatment of tumors, as well as age related macular degeneration. PDT is initiated by introducing a photosensitizer agent into a subject's blood stream. After an appropriate time interval (usually tens of hours), the photosensitizer is activated by shining a visible light, usually a red color laser beam, at the tumor's location.

PDT employs the special ability of some po hyrin and porphyrin-like photosensitizers to accumulate in pathologic cells, and to transfer, upon or subsequent to radiation, absorbed photon energy to naturally occurring oxygen molecules in blood and tissue. Photophysical processes constituting PDT using porphyrins agents are summarized in the energy level diagram shown in Fig. 1. l-WA/1851229.1 In its classical implementation, absorption of one photon of visible wavelength takes a photosensitizer molecule into a short-lived excited state, Si, with energy of 170-190 kJ mol^"1 , which corresponds to an illumination wavelength of about 620 to 690 nm. After a few nanoseconds, the poφhyrin converts into a triplet state, Ti, by an intersystem crossing (ISC) mechanism with energy of 110-130 kJ mol^"1 and a much longer lifetime, on the order of milliseconds. From this triplet state, energy is transferred to omnipresent oxygen molecules by switching them from a triplet ground state, 3∑g, into an excited singlet state, lΔg, which has an excitation energy of 94 kJ mol-1. Once in the excited singlet state, the oxygen presents an extremely active species, which reacts chemically with the surrounding cell material and causes tumor apoptosis.

The use of longer wavelength, near-infrared, light to cause absorption of two photons of longer wavelength light has been developed to treat breast and other cancers. See U.S. Patent Nos. 5,829,448, 5,832,931, 5,998,597, and 6,042,603. This two-photon technique employs a mode-locked Ti:sapphire laser to administer PDT with near-infrared light. In contrast to one-photon PDT, the near-infrared light produced by the Ti:sapphire laser is at a wavelength substantially longer than the characteristic one-photon absorption waveband ofthe photoreactive agent employed. Instead ofthe single photon absorption process involved in a conventional photodynamic reaction, a two photon process may occur upon radiation with a pulse ofthe 700-1300 nm light. Due to its relatively long wavelength, the near-infrared light emitted by a Ti: sapphire laser can penetrate into tissue up to 8 centimeter or more, making it possible to treat tumors that are relatively deep within a subject's body, well below the dermal layer.

For photosensitizer molecules to be particularly efficacious they should selectively accumulate in the tumor tissue. It is known that poφhyrin-based molecules possess this feature. To date, the U.S. Food and Drug Administration has approved at least two porphyrin-based PDT agents: Photofrin®, and

Nerteporfrin® . Photofrin® is a naturally occurring porphyrin, which absorbs light in the visible spectral range (λ<690nm). However, neither of these compounds have significant absorption spectra in the near-infrared region of radiation of 700 to 1300 nm, nor do they exhibit efficient multi-photon absorption. Chemical modification ofthe porphyrin structure, such as to chlorin or bacteriochlorin, to shift the one-photon absorption band to longer wavelengths is limited by the fundamental requirement that the energy ofthe Ti state be higher than the excitation energy of singlet oxygen. Furthermore, such structural modification ofthe porphyrin structure may result in a less stable compound.

Non-porphyrin-based materials may have enhanced TPA cross-sections but typically lack either the ability to generate singlet oxygen, or have either unknown or deleterious interaction properties with biological tissue.

Thus, there is a need of porphyrin-based materials which safely interact with biological tissue and exhibit both a significant two-photon absorption cross-section for radiation in the near-infrared region and the ability to generate singlet oxygen. There is also a need to effectively treat tumors and other manifestations of disease located deep within a subject's body by PDT. Presently known and approved PDT agents require the use of radiation ranging from about 620 nm to 690 nm. This range of radiation typically penetrates most tissues to a depth of no more than a few millimeters.

SUMMARY OF THE INVENTION The invention meets the need to treat physical manifestations of diseases or disorders located within a subject's body with photodynamic therapy by providing porphyrin-based compounds which undergo simultaneous two photon absoφtion upon exposure to radiation easily transmitted by a subject's tissue to produce, in due course, singlet oxygen. The use of radiation in the tissue transmission region permits the treatment of disorders located more than a few millimeters within a subj ect' s body. In one embodiment, the invention relates to poφhyrin-based compounds modified with chromophores to provide enhanced multi-photon absoφtion of radiation in the range of about 700 nm to about 1300 nm. One or more ofthe chromophores may be attached to the poφhyrin ring at any one ofthe 2, 3, 5, 7, 8, 10, 12, 13, 15, 17, 18 or 20 positions, that is, at either one ofthe "beta" or "meso" positions ofthe poφhyrin. The chromophores may be directly attached to the ring structure or there may be a linking structure between the poφhyrin ring and the chromophore. The inventive compounds may also be based on various reduced forms of poφhyrin, such as, for example, chlorin (2,3- dihydropoφhyrin), bacteriochlorin (7,8, 17, 18-tetrahydropoφhyrin), and isobacteriochlorin.

In another general embodiment, the invention is directed to a method for generating singlet oxygen by irradiating a poφhyrin-based compound with multiple photons, generally two photons, of near-infrared radiation. The compound may comprise a poφhyrin-based compound substituted with one or more chromophores at the beta or meso positions, and further substituted with an intersystem crossing enhancing substituent also at the beta or meso positions.

Another general embodiment ofthe invention is further directed to a method of treating a tumor disease or a tumor by contacting an therapeutically effective amount of a poφhyrin-based compound modified with one or more chromophores and at least one intersystem crossing enhancing substituent as set forth herein and irradiating the modified poφhyrin with sufficient radiation to produce singlet oxygen. Preferably, the modified poφhyrin absorbs two photons of radiation which radiation is preferably is in a range of about 700 nm to about 1300 nm.

In another embodiment, the invention relates to a pharmaceutical use of the inventive poφhyrin compound in an amount that is therapeutically effective for the treatment of tumors or other disorders.

In yet another embodiment, the invention is directed to a method of increasing the multi-photon absoφtion cross-section of a poφhyrin-based photosensitizer by attaching at least one TPA-chromophore at the meso or beta positions of a poφhyrin structure ofthe poφhyrin-based photosensitizer, and also attaching at least one intersystem crossing enhancing substituent to meso or beta positions of a poφhyrin structure ofthe poφhyrin-based photosensitizer. These attachments result in an increased multi-photon absoφtion cross-section ofthe poφhyrin-based photosensitizer and significant ability to generate singlet oxygen. The increased multi-photon absoφtion cross-section may be at least about 30 GM units, preferably at least about 50 GM units, and more preferably at least about 70 GM units. The absoφtion cross-section is measured at about the photosensitizer' s maximum wavelength for two-photon absoφtion.

A further embodiment ofthe invention may include a poφhyrin-based compound comprising the following poφhyrin structure:

wherein each R¹ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, - CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; each R² is independently -

L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; each R³ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; or independently R² and R³ are linked by -C₄H₄- to form a six-member ring; M is either two hydrogen atoms or a metal ion; each n is independently an integer ranging from 1 to 20; each G is independently -H, or a Ci to C₂₀ alkyl; Ar comprises one of C₆H₅, C₆H₄X, C₆H X₂, C₆H₂X₃, H j, or C₆X₅, wherein X comprises one of F, Cl, Br, or I; L is a linking moiety between the poφhyrin structure and TPA; TPA is a chromophore moiety; and wherein at least one of R , R or R is a L-TPA moiety, and at least one other of R¹, R² or R³ is not H or a L-TPA moiety.

Preferably, the TPA chromophore moiety may be attached to the poφhyrin ring directly or with a linking group, L. The linking group may be selected from the group consisting of ethenyl, ethynyl, -(CH₂)n- wherein is equal to 1 to 20, ortho-phenyl, meta-phenyl, para-phenyl, -C(=O)-O-, and 4- phenyl-2'-ethenyl.

The TPA-chromophore may be any ofthe following structures:

TPA A

wherein in TPA A, n = 1 to 5, and R comprises one member selected from the group consisting of H, alkyl, alkyloxy, -(OCH₂CH₂)nOG; wherein G is H or alkyl;

TPA B

wherein in TPA B, n = 1 to 5, and R comprises an alkyl;

TPA C

wherein in TPA C, n = 1 to 3, R comprises one member selected from the group consisting of H, CN, alkyl, alkyloxy, and R' comprises one member selected from the group consisting of alkyl, alkyloxyphenyl, phenyl, phenyl- (OCH₂CH₂)nOG; wherein G is H or alkyl;

TPA D wherein in TPA D, n = 1 to 5, and R' comprises one member selected from the group consisting of alkyl, alkyloxyphenyl, phenyl-(OCH CH₂)nOG; wherein G is H or alkyl;

TPA E wherein in TPA E, n = 1 to 5, R comprises one member selected from the group consisting of H, alkyl, and -(OCH₂CH₂)_nOG; wherein G is H or alkyl, and R' comprises alkyl;

TPA F

wherein in TPA F, n = 1 to 3, R comprises one member selected from the group consisting of H, alkyl, (OCH₂CH₂)_nOG; wherein G is H or alkyl, and R' comprises alkyl;

TPA G

wherein in TPA G, R comprises one member selected from the group consisting of H, CN, alkyl, alkyloxy, -(OCH₂CH₂)_nOG, and R' comprises one member selected from the group consisting of H, alkyl, alkyloxy, - (OCH₂CH₂)_nOG; and wherein G is H or alkyl and n = 1 to 6;

TPA H

wherein in TPA H, R comprises one member selected from the group consisting of alkyl, phenyl, alkyloxyphenyl, phenyl[(OCH₂CH₂)nOG]x; wherein G is H or alkyl, n=l to 4, and x=l to 4; wherein in TPA's A to H, alkyl comprises Ci to C₂₀ alkyl moieties; and wherein the TPA may be attached to the poφhyrin ring at the point indicated by the wiggle line (" _'wn-n, "). The poφhyrin structure ofthe poφhyrin-based photosensitizer may be any one of poφhyrin, chlorin, bacteriochlorin, and isobacteriochlorin. The poφhyrin-based photosensitizer may absorb two photons of radiation in the range of about 700 nm to about 1300 nm, preferably the range is about 700 nm to about 1100 nm.

Another embodiment ofthe invention is improving the method of conducting photodynamic therapy which comprises providing a pharmaceutically effective amount of a photoactive compound to within a subject, contacting the photoactive compound with an area to be treated within a subject, and exposing the area to radiation with a wavelength ranging from between about 700 nm to about 1300. The improvement comprises utilizing as the photosensitizer the poφhyrin-based photosensitizer described herein.

Additional features, advantages, and embodiments ofthe invention may be set forth or apparent from consideration ofthe following detailed description, drawings, and claims. Moreover, it is to be understood that both the foregoing summary ofthe invention and the following detailed description are exemplary and intended to provide further explanation without limiting the scope ofthe invention as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are included to provide a further understanding ofthe invention and are incoφorated in and constitute a part of this specification, illustrate preferred embodiments ofthe invention and together with the detailed description serve to explain the principles ofthe invention, hi the drawings:

Figure 1 - Schematic ofthe energy levels for a poφhyrin-based photosensitizer (solid bars) and molecular oxygen (open bars). Figure 2 - Molecular structures of three poφhyrins: Compound I - 5-[4^m-(diphenylamino)-4"-stilbenyl]- 15-[2',6'- dichlorophenyl]-21H, 23H-poφhyrin ("DPASP"); Compound II - 5-phenyl, 15-(2',6'-dichlorophenyl)-21H, 23H-poφhyrin; and

Compound 111 - 5,10, 15,20-tetraphenyl-2 lH,23H-poφhyrin. Figure 3 - Two-photon absoφtion spectra (circles) of 10^"4 M toluene solutions of compounds I, II, and III.

Figure 4 - (a) The ^lΔ_g - ³∑_g luminescence spectra of molecular oxygen in an air-saturated toluene solution of compound I.

(b) The dependence ofthe ^lΔ_g -> ³∑_g luminescence intensity {l_) on the average illumination intensity, P, of molecular oxygen in an air-saturated toluene solution of compound I.

Figure 5 - Poφhyrin design motifs for enhancement of their intrinsic two-photon cross-sections.

Figure 6 - TPA chromophore motifs with enhanced intrinsic two-photon absoφtion. Figure 7 - Linker moieties for attachment of TPA chromophores to poφhyrins.

Figure 8 - Conversion of m-THPC to a new derivatized poφhyrin with enhanced two—photon absoφtion.

Figure 9 - Scheme showing preparative route for (diphenylamino)- substituted aldehydes

Figure 10 - Scheme showing preparative route for α,ω-bis- (diphenylamino)diphenylpolyenes

Figure 11 - Scheme showing preparative route for α,ω-bis- (dialkylamino)diphenylpolyenes Figure 12 - Scheme showing preparative route for α,ω-bis-

(diphenylamino)-PPN-dimer and α,ω-bis-(dialkylamino)-PPN-dimer (PPN = paraphenylene vinylene)

Figure 13 - Scheme showing preparative route for diphenylamino compounds Figure 14 - Scheme showing preparative route for substituted dithienylpolyenes Figure 15 - Scheme showing preparative route for substituted PTN oligomers (PTN = 2,5-dithienylene vinylene)

Figure 16 - Scheme showing preparative route for dithienylpolyene precursors Figure 17 - Scheme showing preparative route for ditliienylpolyenes

Figure 18 - Scheme showing routes to functionalize terminal aryl rings of chromophores for poφhyrin attachment

Figure 19 - Scheme showing routes to functionalize terminal thienyl rings of chromophores for poφhyrin attachment Figure 20 - Molecular structures of two poφhyrins :

Compound IN - 5'-[4'"-(diphenylamino)-4"-stilbenyl-r-ethenyl]- (2,7,12,17-tetra-t-butyl)-21H, 23H-poφhyrin; and

Compound N - 2'-[4'"-(diphenylamino)-4"-stilbenyl-r-ethenyl]- (3,8,13,18-tetra-t-butyl)-21H, 23H-poφhyrin.

DETAILED DESCRIPTION OF THE INVENTION

The energy change associated with photon excitation of a photosensitizer and subsequent energy transfer to produce excited singlet oxygen is represented in Fig. 1. The comparative energy levels for a poφhyrin- based photosensitizer and molecular oxygen are represented by solid and open bars, respectively. The ground, first singlet, ith excited singlet, and lowest triplet states ofthe photosensitizer are labeled S₀(g), Sj(u), S;(g), and Ti, respectively. The symbols in the parenthesises denote the gerade (g) and ngerade (u) symmetry ofthe corresponding states. The ground and the first excited singlet states of molecular oxygen are denoted by ∑_g and Δ_g, respectively.

Upon one-photon excitation ("OPE") the lowest singlet state, Sι(u), of the photosensitizer is populated. Subsequently, the radiationless intersystem- crossing ("ISC") Sι(u) - Ti takes place. This is followed, in turn, by energy transfer (ET) between the photosensitizer and oxygen; Ti + ³∑_g -> S₀(g) + ^λΔ_g (semicircle arrows). The excited singlet oxygen molecules {^lΔ_s) are detected by their ^lΔ_g -^ ³∑_g luminescence at -1270 nm. Upon two-photon excitation ("TPE"), a transition occurs to one ofthe higher excited states S;(g). In the TPE case the energy of each excitation photon is less than the single excitation photon used in OPE, and the energy ofthe TPE excitation photon falls into the tissue transparency window. By the internal conversion ("IC") process, the S;(u) state ofthe photosensitizer is ultimately populated from the Si(g) state, and the remaining steps in the process are the same as in the OPE case described above. As presented in Fig. 3, the two-photon absoφtion spectra of compounds

I, II, and III could not be measured at wavelengths shorter than about 350 nm because the energy ofthe excitation photon approaches that ofthe first excited singlet state Si, and efficient one-photon absoφtion masks TPA.

Fig. 3 has a horizontal (wavelength) axis corresponding to the transition wavelength, that is, it is half the laser excitation wavelength. The vertical axis shows the TPA cross-sections in Gδppert-Mayer units ("GM", 1 GM = 10^"50 cm⁴ s photon^"1 molecule^"1); note the differing scales on the three graphs. The laser system comprised a phase-locked Ti:sapphire regenerative amplifier (Model CPA-1000 made by Clark-MXR, Inc.(Dexter, MI)), which was operated at 1 kHz repetition rate and produced 150-fs pulses at 0.6 mJ energy per pulse. These pulses were parametrically down-converted in an optical parametric amplifier ("OP A"), (TOP AS made by Quantronix (East Setanket, NY)) which yielded 100-fs pulses in the range from 1170 to 1640 nm. TPA spectra were obtained by tuning the OPA with subsequent second harmonic generation and registration ofthe poφhyrin fluorescence. Absolute TPA cross-sections were measured by comparing fluorescence intensity under one- and two-photon excitation (see Drobizhev, M., et al. in Chem. Phys. Lett. Vol. 334, pages 76-82 (2001) and references therein for details). Black solid lines represent the appropriately scaled one-photon absoφtion. All three molecules reveal a characteristic strong Soret band near 415 nm, accompanied by four relatively weak bands in the visible (not shown). As illustrated in Fig. 4(a), the ^lΔ_g - ³∑_g luminescence spectra of molecular oxygen in an air-saturated toluene solution of compound I after both one- and two-photon excitation were measured. The one-photon excitation is represented by the dashed line and the two-photon excitation is shown by the solid line. Both spectra are normalized to unity. For one photon excitation, second harmonics of a Ti: sapphire regenerative amplifier, 390 nm, with an average power intensity of 0.5 W/cm² were used, while for two-photon excitation, the output of a Ti:sapphire regenerative amplifier, 780 nm, with an average power intensity of 15 W/cm² was used. In both cases, the laser beam was slightly focused into a 1-cm cell to give a cylindrical irradiated volume of ~1.5 mm diameter. The singlet oxygen luminescence spectrum was measured with a nitrogen-cooled Ge detector coupled with Jobin-Yvon monochromator and a lock-in amplifier.

Fig. 4(b) graphs the dependence ofthe ^lΔ_s - ³∑_g luminescence intensity {l_) on the average illumination intensity, P, of molecular oxygen in an air-saturated toluene solution of compound I after two-photon excitation. The raw experimental data are shown by black squares, with the best power-law fit, l_ = aPⁿ with n = 2.1 + 0.1 shown by the dotted line. These results confirm the two- photon nature of singlet oxygen production. The present invention is directed to a method of increasing the multi- photon absoφtion cross-section of a poφhyrin-based photosensitizer by attaching at least one TPA-chromophore at the meso or beta positions of a poφhyrin structure ofthe poφhyrin-based photosensitizer, and attaching at least one intersystem crossing enhancing substituent to meso or beta positions of a poφhyrin structure of the poφhyrin-based photosensitizer.

This modification ofthe poφhyrin photosensitizer increases the multi- photon absoφtion cross-section ofthe poφhyrin-based photosensitizer to at least about 30 GM units at about its maximum wavelength for two-photon absoφtion. The TPA-chromophore may be selected from the following structures: the following structures:

TPA A

TPA B

wherein in TPA B, n = 1 to 5, and R comprises an alkyl;

TPA C

TPA D

wherein in TPA D, n = 1 to 5, and R' comprises one member selected from the group consisting of alkyl, alkyloxyphenyl, phenyl-(OCH₂CH₂)nOG; wherein G is H or alkyl;

TPA E wherein in TPA E, n = 1 to 5, R comprises one member selected from the group consisting of H, alkyl, and -(OCH₂CH )_nOG; wherein G is H or alkyl, and R' comprises alkyl;

TPA F

TPA G

wherein in TPA G, R comprises one member selected from the group consisting of H, CN, alkyl, alkyloxy, -(OCH₂CH₂)_nOG, and ' comprises one member selected from the group consisting of H, alkyl, alkyloxy, - (OCH₂CH₂)_nOG; and wherein G is H or alkyl and n = 1 to 6;

TPA H

wherein in TPA H, R comprises one member selected from the group consisting of alkyl, phenyl, alkyloxyphenyl, phenyl[(OCH₂CH₂)_nOG]_x; wherein G is H or alkyl, n=l to 4, and x=l to 4. For TPA's A to H, alkyl comprises d to C₂₀ alkyl moieties; and the TPA is attached to the poφhyrin ring at the point indicated by the wiggle line (" 'wvα^, ").

The TPA-chromophore may be attached directly to the poφhyrin ring or via a linking group. The linking group may be any one of ethenyl, ethynyl, - (CH )n- wherein n is equal to 1 to 20, ortho-phenyl, meta-phenyl, para-phenyl, - C(=O)-O-, and 4-phenyl-2'-ethenyl.

Without limiting the invention, the present theory regarding the generation of singlet oxygen, as illustrated in Fig. 1, involves an intersystem crossing step. A photosensitizer' s ability to generate singlet oxygen appears to be significantly influenced by the ease at which it undergoes the intersystem crossing step. This ability may be enhanced by the addition of ISC enhancing substituents to the poφhyrin ring.

These intersystem crossing enhancing substituents include the group consisting of C₆H₅, CδELX, C₆H₃X₂, C₆H₂X₃, CβHXt, or C₆X₅, wherein X may be one of F, Cl, Br, or I. The halogens may be substituted onto the phenyl ring in any combination of halogens and ring positions.

The poφhyrin ring is the preferred ring structure for the photosensitizer but reduced poφhyrin forms, such as, chlorin, bacteriochlorin, and isobacteriochlorin, may be utilized as the basic ring structure for the photosensitizer.

Preferably, the poφhyrin-based photosensitizer absorbs two photons of radiation in the range of about 700 nm to about 1300 nm, more preferably the absorbed radiation is in a range of about 700 nm to about 1100 nm.

The poφhyrin-based photosensitizer may be based on the following comprising the following poφhyrin structure:

wherein each R¹ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, - CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; each R² is independently - L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; each R³ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂, - (CH₂O)n-G, -t-Butyl, or -C(O)OG; or independently R² and R³ are linked by - C₄H₄- to form a six-member ring; M is either two hydrogen atoms or a metal ion; each n is independently an integer ranging from 1 to 20; and each G is independently -H, or a Ci to C₂₀ alkyl.

Ar comprises one of C₆H₅, C₆H₄X, C₆H₃X₂, C₆H₂X₃, C₆HX₄, or C₆X₅, wherein X comprises one of F, Cl, Br, or I; L is a linking moiety between the poφhyrin structure and TPA; and TPA is a chromophore moiety as described above. At least one ofthe R¹, R² or R³ poφhyrin-ring substituents is a L-TPA moiety, while at least one other of R¹, R² or R³ is not H or a L-TPA moiety. Preferably, one of R¹, R² or R³ is an Ar group.

The TPA moiety may be attached to the poφhyrin ring directly or with a linking group, L. The linking group may be one of ethenyl, ethynyl, -(CH₂)n- wherein n is equal to 1 to 20, ortho-phenyl, meta-phenyl, para-phenyl, -C(=O)- O-, and 4-phenyl-2'-ethenyl. Most preferably, the compound may be 5-[4'"- (diphenylamino)-4"-stilbenyl]-15-[2',6'-dichlorophenyl]-21H, 23H-pθφhyrin.

Additionally, M may be any metal atom. Preferred are metals which are biologically acceptable to a subject's body without deleterious health effects. Alternatively, M may be two hydrogen atoms.

The compounds described herein may be advantageously utilized as PDT agents in the treatment of tumors or other physical manifestations of a disease or condition. Such a use would be an improvement over the PDT treatments and agents presently known, by allowing the effective use of treatment radiation with a wavelength ranging from between about 700 nm to about 1300 nm, radiation which easily passes through living tissue to produce singlet oxygen. Preferably, the PDT agent absorbs two photons of radiation in a range of about 700 nm to about 1100 nm. An improved treatment method calling for the pharmaceutical use ofthe inventive poφhyrin compound in an amount that is therapeutically effective for the treatment of tumors or other disorders with an improved two photon absoφtion cross-section at about its maxima for two-photon absoφtion is another application ofthe compounds. A therapeutically effective amount of the compound would range from about 0.1 mg to about 10 mg per kg of body weight ofthe subject. The chromophores and poφhyrin with the chromophores attached may be prepared by following the general synthetic procedures outlined in Figs. 9 to 19. Specifically, preparative routes for ammo-containing chromophores are outlined in Figs. 9 to 13, while thienyl-containing chromophore preparative routes are set forth in Figs. 14 to 17. Methods of functionalizing the terminal aryl or thienyl group ofthe chromophores for subsequent attachment to the poφhyrin ring at the desired position or positions by means of general coupling reactions are set forth in Figs. 18 to 19.

EXPERIMENTAL DETAILS

The solvents and reagents used were obtained from Fisher Scientific and Aldrich Chemical Companies and used as received.

Compound II was prepared via the same preparative route as set forth for compound I below. Compounds I and II are purified by flash chromatography on silica using 20-40% dichloromethane/hexane gradient. Molecular structures were confirmed by spectroscopic analysis. Compound III was purchased from Aldrich and used as received.

EXAMPLE 1 - 5-[4'"-(diphenylamino)-4"-stilbenyl]- 15-[2',6'-dichlorophenyl]- 21H, 23H-poφhyrin (Compound I)

Trifluoroacetic acid (17.2 μL, 0.22 mmol) was added to a mixture of 10- (2,6~dichlorophenyl)bilane (100 mg, 0.15 mmol) and 4'- diphenylaminostilbene-4"-carbaldehyde (83.6 mg, 0.15 mmol) in dichloromethane (23 mL) at RT under an argon atmosphere in the absence of light. After stirring for 1 hour, 2,3-dichloro-4,5-dicyano-l,4-benzoquinone (227 mg, 0.67 mmol) was added and the resulting mixture stirred for an additional 1 hour. Triethylamine (137 [μL, 0.98 mmol) was then added, and the mixture stirred for 5 minutes, and then filtered through a short column of silica using hexanes and dichloroethane to separate out the crude poφhyrin. After solvent removal, the title poφhyrin was obtained by flash chromatography (6.5 cm diameter, 5.5 cm silica height, eluting with dichloromethane/hexanes, 1:4 to 2:3 gradient) as apuφle solid (95 mg, 53%)

EXAMPLE 2 - Precursors for Compounds TV and V - 2-Formyl-3,8,13,18-tetra- t-butylpoφhyrin ("TTBP") and 5-formyl-2,7,12,17-tetra-t-butylpθφhyrin

A mixture of dimethylformamide (1.30 mL, 17 mmol) and phosphorus oxychloride (1.30 mL, 14 mmol) was stirred for 10 minutes at 40 °C. Ni(II)- 2,7,12,17-tetra-t-butylpoφhyrin ("TTBP") (1.27 g, 2.15 mmol) and dichloroethane (DCE) (70 mL) were then added, and the resulting mixture stirred at 90 °C for 12 hours. The reaction mixture was then poured into a solution of sodium acetate (10.4g, 77 mmol) in water (5 mL) and heated to reflux for 1 hour. After cooling to RT, DCE and water (150 mL each) were added and the layers separated. The aqueous layer was extracted with DCE (3 x 150 mL). The combined DCE layers were then washed with Na₂CO₃ (1 x 50 mL) and dried with MgSO₄. The filtered solution was concentrated under reduced pressure to ca. 100 mL and preabsorbed onto silica gel (ca. lOg). The crude products were then isolated by flash column chromatography (silica, DCE/hexanes, 20%-70% gradient). The meso isomer (5-formyl) eluted just after unreacted starting material [Ni(II) TTBP] and was isolated as a puφle solid (168 mg, 12.6%). The beta isomer (2-formyl) eluted next, and was isolated as a green solid (532 mg, 40.0%).

The beta (2-formyl) isomer (350 mg) was demetallated by dissolving in the minimum amount of sulfuric acid (80 mL) at RT for 1 minute. The acid solution was then poured into an ice/water mixture (1 Kg/200 mL), neutralized with the minimum amount of NaOH to cause the product to precipitate completely, filtered and washed with warm water. The wet material was then dissolved in a minimum volume of DCE and dried with MgSO₄. The solvent was removed under reduced pressure, and the product further dried in vacuo (310 mg, 97%). The same procedure was employed for the demetallation ofthe meso (5-formyl) isomer.

EXAMPLE 3A - Attachment of TPA Group, 2-[4"-(diphenylamino)-4'- stilbenyl]-l-ethenyl), to meso position of 5-formyl-TTBP to form Compound IN Potassium tert-butoxide (0.2 mL, 1 M solution in THF) was added to a mixture of meso-formyl-TTBP (7.2 mg, 0.013 mmol) and 4'-(diphenylamino)-4- stilbenyl-methyltributylphosphonium bromide (90 mg, 0.14 mmol) in anhydrous THF at 0 °C. The mixture was stirred at RT for 20 minutes, then refluxed for 48 hours. After cooling, the product was extracted with methylene chloride, and the resulting solution washed with brine and dried with MgSO₄. After the solvent was removed by rotovaporization, the product was purified by basic alumina column chromatography (CH₂Cl₂/hexane, 1:1). The product was further purified by recrystallization. from MeOH/CH Cl₂ (10 mg, 86%).

1H ΝMR δ (-3.85) (s, 2H, ΝH), 2.14-2.41 (s, 36 H, CH3),6.97-7.86 (m, 22H, Ar-H and vinyl H), 10.42, 10.45, 10.61 (3 s, meso CH), 9.12, 9.20, 9.22, 10.74 (4s, 4H, pyrrolic-H); 13C ΝMR 34.01, 34.14, 34.52, 34.73, 34.92, 37.05, 37.72, 77.00, 77.43, 77.85, 102.97, 103.10, 105.13, 105.38, 125.78, 127.02, 131.30, 194.55; MALDI-TOF, m/z: 920.47, 906.36, 848.29, 665.26.

EXAMPLE 3B - Attachment of TPA Group, 2-[4"-(diphenylamino)-4'- stilbenyl]-l -ethenyl), to beta position of 2-formyl TTBP to form Compound N

Potassium tert-butoxide (0.15 mL, 1 M solution in THF) was added to a mixture of beta-formyl-TTBP (12.5 mg, 0.022 mmol) and [4'-(diphenylamino)- 4-stilbenyl]-methyltributylphosphonium bromide (50 mg, 0.078 mmol) in anhydrous THF at 0 °C. The work-up and isolation ofthe desired product was identical to that described above for the meso isomer (12 mg, 60%). 1H NMR δ (-2.93) (bd. s, 2H, NH), 2.16-2.27 (s, 36 H, CH3), 7.02-7.85 (m, 22H, Ar-H and vinyl H), 9.01, 9,12, 9.22 (3s, 3H, pyrrolic-H), 9.33, 10.20, 10.42 (3s, 4H, meso-H); 13C NMR 33.94, 34.15, 35.01, 76.98, 77.40,77.83, 123.51, 123.95..124.98, 127.01, 127.39,127.63,127.87,129.72 ppm; MALDI- TOF, m/z: 938.60, 906.61, 681.46, 665.47.

EXAMPLE 4 - Bis-(4',4"-diphenylamino)stilbene

A 500 mL 3-neck round bottom flask equipped with a reflux condenser, magnetic stir bar, nitrogen inlet and rubber septum was charged with anhydrous THF (150 mL). The flask was cooled to 0 °C, and titanium tetrachloride (7.36 mL, 0.0388 mol, 2 eq.) was added dropwise by syringe. Zinc (5.07g, 0.0776 mol, 4 eq.) was then added in small portions to the emulsion, and the resulting mixture was refluxed for 45 minutes. The reaction mixture was cooled to 0 °C and a solution of 4-formyltriphenylamine (5.3g, 0.0194 mol, 1 eq.) in anhydrous THF and pyridine (5 mL) was added dropwise from an addition funnel. The mixture was then refluxed, and the progress monitored by TLC (dichloro ethane) until completion. The mixture was then cooled and poured into water (80 mL). The resulting emulsion was stirred for 20 minutes and then partitioned in a separatory funnel. The aqueous layer was extracted with dichloromethane (100 mL), and the combined organic layers were washed with water (3 x 80 mL). The solution was then dried with MgSO and the solvent removed by rotovaporization. The resulting solid was recrystallized from ethanol to yield a yellow solid (4.45g, 90%),1H MR δ 6.47-7.45 (m, 30H, Ar- H and CH=CH), 13C NMR122.7, 123.3, 123.6, 124.1, 124.8, 126.9, 127.5, 129.6, 130.2, 132.3, 147.4, 147.9 ppm.

EXAMPLE 5 - General Procedure for Oxopropenylation of Diphenylaminophenylaldehydyes A 3-neck round bottom flask equipped with a magnetic stir bar, CaC12 drying tube, nitrogen inlet and stopper was charged with the selected aldehyde in anhydrous THF (1 eq.). Tributyl-(l,3-dioxolan-2-ylmethyl)phosphonium bromide in THF (1.1 eq.) was added, then potassium tert-butoxide (1.4 eq.) was added in small portions. The reaction was stirred at RT until completion as indicated by TLC (1: 1 dichloromethane :hexane). After completion, aqueous HCI (2M, 50 mL) was added dropwise, and the resulting bilayer separated. The aqueous layer was extracted with dichloromethane (3 x 75 mL), and the combined organic solution was washed with water (3 x 75 mL) and dried with MgSO₄. The solvent was removed by roto vaporization and the resulting solid purified by silica gel chromatography (1:1 dichloromethane:hexane).

The following compounds are provided as examples of this general extension procedure:

3-[4'-(diphenylamino)phenyl]-2-propenal (72%), m.p. 109-11 °C, λmax 395 nm (amax 35,179), 1HNMR δ 6.62-6.55 (dd, 1 H, Jab=15.8, Jbc=7.8 Hz,

=CHCHO), 6.99-7.41 (m, 15H, Ar-H, and 1 vinyl H), 9.60-9.62 (d, Jcb=7.8 Hz, CHO), ¹³C NMR 121.4,124.8, 126. 1, 126.4, 127.1, 129.9, 130.2, 146.9, 151.2, 153.0, 194.1 ppm.

5-[4'-(diphenylamino)phenyl]-2,4-pentadienal (73%), m.p. 137-9 °C, λmax. 413 nm (amax 43,904), 1H NMR δ 6.18-6.26 (dd, 1H, Jab 15 Hz, Jb„ 8 Hz, =CHCHO), 6.82-7.36 (m, 17H, Ar-H and 1 vinyl H), 9.60 (d, 1 H, Jbc 8 Hz, CHO), 13C NMR 122.3,124.3,124.4, 125,2, 125.7, 129.0, 129.8, 130.8, 142.7, 147.3, 149.7, 153.1, 193.9 ppm.

EXAMPLE 6 - General procedure for the synthesis of α,ω-bis- (diphenylamino)diphenylpolyenes

A 3-neck round bottom flask equipped with a magnetic stir bar, CaC12 drying tube, nitrogen inlet and stopper was charged with the appropriate aldehyde (1 eq.) in anhydrous THF. A bis-Wittig salt (0.5 eq.) in THF is then added: (E)-but-2-en-l,4-diyl-bis-(tributylphosphonium) dichloride for polyenes with an odd number of double bonds, and (E,E)-hexa-2,4-dien-l,6-diyl-bis- (tributylphosphonium) dibromide for polyenes with an even number of double bonds. Potassium tert-butoxide (2.2 eq.) is then added in small portions, and the reaction mixture is stirred at RT until completion as indicated by TLC

(dichloromethane). The solvent is removed by rotovaporization, and the impure product is purified by silica gel column chromatography (dichloromethane).

The following compounds are provided as examples of this general synthesis procedure:

Bis-(4',4"-diphenylamino)-l,6-diphenyl-l,3,5-hexatriene (82%), λmax 420 nm (amax, 64,630), 1H NMR δ 6.4-6.8 (m, 6H, vinyl), 7.00-7.23 (m, 28 H, Ar-H), 13C NMR 123.4,123.9, 124.9, 127.6, 128.2, 129.7, 132.1, 133.4, 147.5, 147.9 ppm.

Bis-(4',4"-diphenylamino)-l,8-diphenyl-l,3,5,7-octatetraene (81%), λmax 434 nm amax (35,588), 1H NMR δ 6.30-6-89 (m, 8H, vinyl), 7.02-7.25 (m, 28H, Ar-H), 13C MR 123.5, 123.9, 124.9, 127.6, 128.1, 129.6, 132.2, 133.2, 133.7, 147.5, 147.9 ppm.

EXAMPLE 7 - 4-(Diphenylamino)-4'-formylstilbene

Potassium tert-butoxide (0.044 mol, 44mL 1M solution in THF) was added to a mixture of terephthalaldehyde mono-(diethyl acetal) (Sigma- Aldrich) (9.98g, 0.048 mol) and 4-(diphenylamino)benzyltributylphosphonium bromide (21.6g, 0.040 mol) in anhydrous THF (200 mL) in a 500 mL round bottom flask equipped with a reflux condenser, addition funnel and nitrogen inlet. After the addition was complete, the mixture was refluxed for 2 hours, and then cooled to RT. Aqueous HCI (20 mL, 3M) was then added and the mixture was stirred an additional hour. The product was extracted with methylene chloride (3 x 100 mL) and the combined organic solution dried with MgSO₄. The solvent was removed by rotovaporization and the crude product purified by silica gel column chromatography (1:1 hexane methylene chloride) yielding the pure product as a bright yellow solid (13g, 87%), 1H NMR δ 7.02-7.85 (m, 18 Ar-H, 2 vinyl H), 9.96 (s, 1H, CHO).

EXAMPLE 8 - 4-(Diphenylamino)-4'-hydroxymethylstilbene

A 500 mL round bottom flask was charged with LAH (0.228g, 0.006 mol) in THF (1 M solution), and compound 17 (4.38g, 0.0117 mol) dissolved in anhydrous THF (100 mL) was added dropwise via an addition funnel. The solution was stirred at RT for 4 hours, cooled to 0 °C, and then water was added to quench the unreacted LAH. The reaction was neutralized with aqueous HCI (4 mL, 3M), and stirred an additional 1 hour. The product was extracted with methylene chloride (3 x 100 mL), and the combined organic product was washed with water (3 x 100 mL) and dried with MgSO₄. After solvent removal by rotovaporization, the product was purified by silica gel column chromatography (4:1 benzene:ethyl acetate) yielding 4-(Diphenylamino)-4'- hydroxymethylstilbene as a yellow solid (3.3g, 75%), 1H NMR δ 4.68 (s, 2H, CH2),7.04-7.45 (m, 18 Ar-H, 2 vinyl H).

EXAMPLE 9 - [4-(Diphenylamino)-4"-stilbenyl]methyltributylphosphonium bromide

4-(Diphenylamino)-4'-hydroxymethylstilbene (l.Og, 0.00265 mol) was dissolved in anhydrous THF (10 mL) and added to a mixture of phosphorus tribromide (0.41g, 0.0015 mol) in anhydrous THF (10 mL) under a nitrogen atmosphere. The resulting mixture was stirred at 0 °C for 2 hours, and then for an additional 16 hours at RT. The product was extracted with methylene chloride, dried with MgSO₄ filtered and the solvent removed by rotovaporization. The product was purified by silica gel column chromatography (CH₂C1₂), dissolved in toluene and reacted with tributylphosphine (0.54g, 0.00265 mol). The resulting solution was refluxed for 72 hours and the product obtained by solvent removal as a glassy solid. The product was dissolved in THF to be used directly in the preparation of IN and V (L 18g₅ 69%), 1H ΝMR δ 0.91 (t, 9H, CH3), 1.47 (m, 12H, CH2), 2.41 (m, 6H, CH2), 4.23 (m, 2H, CH2), 7,0-7.67 (m, 18 Ar-H, 2 vinyl H).

EXAMPLE 10 - Preparation of Precursors for Thienyl-containing Chromophores

2-Butylthiothiophene (44)

A solution of n-butyllithium ( 1.6 M in hexane, 0.304 mol, 189.8 mL was added dropwise to a solution of thiophene (Aldrich) ( 24.3 g, 0.289 mol ) and TMEDA ( 45.8 mL, 0.304 mol ) in THF ( 200 mL ) at room temperature. The resulting mixture was refluxed for 0.5 hours, cooled in an ice-bath, and powdered sulfur ( 9.7 g, 0.304 mol ) added carefully. After the resulting mixture had become clear, iodobutane ( 37.9 mL, 0.333 mol ) was added dropwise. The product mixture was then stirred at room temperature overnight, poured into cold water and extracted with ethyl ether ( 3 X 150 mL). The combined extracts were washed with saturated brine and dried (MgSO₄). After removal ofthe drying agent by filtration, the solvent was removed using a rotary evaporator. The product was obtained as colorless liquid by vacuum distillation (36.2 g, 73%),. b.p. 88-90 °C (1 Torr); 1H ΝMR δ: 0. 89 (t, 3 H, J=7.3 Hz CH₃), 1.4(m, 2 H, CH₃-CH₂),1.58 (m, 2 H, CH₃-CH₂-CH₂), 2.78 (t, 2 H, J=7 Hz, CH₂-S), 6.95 (dd, 1 H J=5.3 Hz, J=3.6Hz, aromatic H), 7.08 (d, 1 H, J+3.6 Hz, aromatic H), 7.3 (d, 1 H, J+5.3 Hz, aromatic H); _λmax /nm (_εmax/dm³ mol^"1 cm^"1) 274 (5500); HRMS (El) calc. for C₈Hι₂S₂ 172.0375, found 172.0380.

5-Butylthiothiophene-2-carb aldehyde (45)

POCl₃ ( 1.7 mL, 0.019 mol ) was added dropwise to a solution of 44 ( 2g, 0.116 mol) and dry DMF ( 1.8 mL, 0.023 mol ) in l,2-dichloroethane(75 mL) at 0°C. The resultant mixture was refluxed for 2 hours, poured into ice water and neutralized by addition of 6 M sodium carbonate solution. The reaction mixture was extracted with methylene chloride ( 3 X 100 mL ). The combined extracts were washed with saturated brine and dried (MgSO₄). After removal ofthe drying agent by filtration, methylene chloride was removed using a rotary evaporator. The residue was purified by column chromatography over silica gel, eluting with CH2C12. The product was obtained as orange liquid (2.2 g, 95%);1H NMR δ: 0.94 (t, 3 H, J=7.3 Hz, CH₃), 1.46 (m, 2 H, CH₃- CH₂), 1.7 (m, 2 H, CH₃-CH₂-CH₂), 3.01 (t, 2 H, J=7.3 Hz, CH₂-S), 7 (d, 1 H, J- 3.8 Hz, aromatic H), 7.61 (d, 1 H, J=3.8 Hz, aromatic H), 9.76 (s, 1 H, CHO); _λraax /nm _max/dm³ mof ¹) 42.5 (11 700); HRMS(EI⁺) calculated for C₉Hι₂OS₂ 200.0320, found 200.0329. The product was used without further purification in the preparation of 46.

2-Hydroxymethyl-5-butylthiothiophene (46)

A solution of NaBH₄ (5.3g, 0.141 mol) in methanol (29.0 g) and sodium hydroxide (20%, 56.0 mL) was added dropwise to a solution of 45 (56.47 g, 0.282 mol) in THF (75 mL) at room temperature. The mixture was stirred for 2 hours, extracted with ethyl ether 3 X 150 mL ), the combined extracts washed with brine and dried (MgSO₄). After solvent removal, the crude product 46 was obtained as an oil (55.4 g, 97%); ^!H NMR δ: 0.88 (t, 3 H, J=7.3 Hz, CH₃), 1.39 (m, 2 H, CH₃-CH₂), 1.57 (m, 2 H, CH₃-CH₂-CH2),1.88 (s, 1 H OH), 2.76 (t, 2 H, J=7.3 Hz, CH₂-S), 4.73 (s, 2 H, CH₂-OH), 6.83 (d, 1 H, J=3.5 Hz, aromatic H), 6.94 (d, 1 H, J=3.5 Hz, aromatic H); λ_max /nm (ε_max/dm mol^" cm^" ) 275.5 (3600); LRMS(EI⁺) calc. for C₉Hι₄OS₂ 202.1, found 202.1. The product was used without further purification in the preparation of 47.

2-Bromomethyl-5-butylthiothiophene (47)

A solution of crude 46 ( 55.4 g, 0.274 mol ) in anhydrous ethyl ether (70 mL was added dropwise to a solution of phosphorus tribromide ( 12.9 mL, 0.137 mol ) in anhydrous ethyl ether ( 300 mL ) at 0 °C. After addition was complete, the mixture was allowed to warm slowly to room temperature and stiffed for 4 hours under nitrogen. The mixture was then extracted with ethyl ether ( 3 X 150 mL ). The combined organic solution was washed with brine and dried (MgSO ). MgSO₄ was removed by filtration and the crude product was carried to the next step without the removal of solvent to prevent product decomposition.

(5-Butylthio-2-thienyl)tributyl methyl phosphonium bromide (48)

Tributylphosphine ( 68.5 mL, 0.274 mol ) was added to a solution of crude 47 in ethyl ether at room temperature. The mixture was stirred under nitrogen for 48 hours. The white solid product was obtained by filtration ( 93 g, 73%) and used without further purification, m.p. 102-104°C; 1H NMR δ: 0.91 (m, 12 H CH₃), 1.46 (m, 16 H, CH₃-CH2), 2.42 (m, 6 H, P-CH₂-CH₂), 2.74 (m, 2 H S-CH₂), 4.55 (s, 1 CH₂-P), 4.6 (s, 1 H, CH₂-P), 6.96 (d, 1 H, J=3.5 Hz, aromatic H), 7.22(d, 1 H, J=3.5 Hz, aromatic H); λ_max /nm (ε_max/dm³ modern^"1) 283.5 (870) (Anal. Calc. for C₂ιH₄₀BrPSO₂: C, 53.95; H, 8.62. Found: C, 54.03; 8.61)

2-(3'-Iodopropoxy) tetrahydro-2H-pyran (59)

2-(3'-Chloropropoxy) tetrahydro-2H-pyran (Aldrich) (9 g, 0.05 mol) was added to a solution of sodium iodide (37.8 g, 0.25 mol) in acetone (250 mL) with stirring. The resultant slurry was refluxed for 48 hours. The mixture was cooled, evaporated to a solid mass and transferred to the top of a short column of neutral alumina. The column was washed with CH C1₂ (1 L) and CH₂CI₂ was removed using a rotary evaporator. The product was obtained as yellow liquid (12.6 g, 93%); 1HNMR δ : 1.62-1.86 (m, 6 H, ring CH₂), 2.07 (m, 2 H, CH₂-CH₂-I), 3.27 (m, 2 H, CH₂-I), 3.3 8-3.54 (m, 2 H, CH₂-O), 3.74-3.89 (m, 2 H ring O- CH₂), 4.58 (t, 1 H, J=3.2 Hz, ring CH); /nm (ε_max/dm³ mol^" ¹) 267 (1500); HRMS (EI⁺) calc. for C₈Hι₅IO₂ 269.0049, found 269.0039.

2-(3 '-Hydroxypropyhhio)thiophene (60) A solution of n-butyllithium 1.6 M in hexane, 0.045 mol, 28.2 mL ) was added dropwise to a solution of thiophene ( 3.62g , 0.043 mol ) and TMEDA ( 5.22g, 0.045 mol) in THF (200 mL ) at room temperature. The resulting mixture was refluxed for 0.5 .hours, cooled in an ice-bath and powdered sulfur ( 1.44g, 0.045 mol ) added carefully with stirring. After the resulting mixture had become clear, 59 ( 12.55g, 0.049 mol ) was added dropwise. The product mixture was then stirred at room temperature overnight, poured into cold water and extracted with ethyl ether (3 x 150 mL). The combined extracts were washed with brine, dried (MgSO ) and solvent was evaporated. The product was obtained as colorless liquid by vacuum distillation. ( 4.8g, 64% ), b.p. 138-140 °C ( I Torr ); 1HNMR δ: 1.58 (s, 1 H, OH), 1.86 (m, 2 H, CH₂-CH₂-OH), 2.9 (t, 2 H, JX.1 Hz, S-CH₂), 3.76 (t, 2 H, J=6.1 Hz, CH₂-OH), 6.97 (dd, 1 H, J=5.3 Hz, J=3.2 Hz, aromatic H), 7.12 (d, 1 H, J=3.2 Hz, aromatic H), 7.34 (d, 1 H, J=5.3 Hz, aromatic H); λ_max /nm (ε_max/dm³ moi^cm^"1) 272.5 (3900); HRMS (EI⁺) calc. for C₇Hι₀OS₂ 174.0173, found 174.0173.

2-(3'-Acetoxypropylthio)thiophene (61)

Acetic anhydride ( 42.9 mL, 0.455 mol ) was added to a solution of 60 ( 13g, 0.075 mol ) in pyridine (57.3 mL, 0.71 mol ) at room temperature. The mixture was stiffed overnight, and then water (100 mL) was added dropwise with an ice-bath cooling. The resultant mixture was stirred for 2 hours and extracted with ethyl ether (3 X 100 mL). The organic layer was washed with water ( 3 X 200 mL ) to remove pyridine. The combined extracts were washed with brine, dried ( MgSO ) and solvent was evaporated. The product was obtained as yellow liquid (14.5 g, 84%); ^!H NMR δ: 1.92 (m, 2 H, CH₂- CH₂-OCOCH₃), 2.03 (s, 3 H, CH₃), 2.84 (t, 2 H, J=7.2 Hz, S-CH₂), 4.16 (t, 2 H, J=6.3 Hz, CH₂-OCOCH₃), 6.98 (dd, 1 H, J=5.2 Hz, J-3.5 Hz, aromatic H), 7.13 (d, 1 H, J=3.5 Hz, aromatic H), 7.35 (d, 1 H, J=5.2 Hz, aromatic H); λ_max /nm (ε_max/dm³ of ¹) 272 (3700). HRMS (EI⁺) calc. for C₉Hι₂O₂S₂ 216.0275, found 216.0279. The product was used directly without further purification in the preparation of 62.

5-(3 '-Acetoxypropylthio)thiophene-2-carbaldehyde (62) POCl₃ (3.4 mL, 0.036 mol ) was added dropwise to a solution of 61 (4.9g, 0.023 mol ) and dry DMF (3.5 mL, 0.045 mol ) in 1,2-dichloroethane (100 mL) at 0 °C. The resultant mixture was refluxed for 2 hours, poured into ice water and neutralized by addition of 6 M sodium carbonate. The reaction mixture was extracted with methylene chloride ( 3 X 100 mL ). The combined extracts were washed with saturated brine, dried (MgSO₄). After filtration, methylene chloride was removed using a rotary evaporator. The residue was purified by column chromatography over silica gel, eluting with 10% ethyl acetate in methylene chloride. The product was obtained as orange liquid (4.5g, 82%) 1H NMR δ: 2 (m, 2 H, CH₂-CH₂-OCOCH₃), 2.03 (s, 3 H, CH₃), 3.04 (t, 2 H, J=7.2 Hz, S-CH₂), 4.15 (t, 2 H, J=6.2 Hz, CH₂-OCOCH₃), 7.04 (d, 1 H, J=3.9 Hz, aromatic H), 7.59 (d, 1 H, J 3.9 Hz, aromatic H), 9.75 (s, 1 H, CHO); /nm (ε_max/dm³ mol^"1 cm^"1) 339 (14 200); HRMS (EI⁺) calc. for Cι₀Hι₂O₃S₂ 244.0231, found 244.0228. The product was used without further purification in preparation of 63.

3-( 5'-Hydroxypropylthio-2'-thienyl)-2-propenal (63)

A solution of potassium tert-butoxide (1 M in hexanes, 0.047 mol, 47 mL) was added dropwise to a solution of 62 (7.63g, 0.031 mol) and (1,3- dioxalane-2-ylmethyl)tributylphosphonium bromide (1 M in THF, 0.03 8 mol, 37.5 mL ) in dry THF ( 150 mL) at room temperature. The resulting mixture was stirred overnight, and then poured into water. The product and tributylphosphine oxide were extracted with ethyl ether ( 3 X 150 mL ), and the combined extracts were washed with saturated brine, and dried (MgSO₄). After filtration, ether was removed by rotary evaporator and the residue dissolved in THF (100 mL ). Aqueous HCI ( 3 M, 120 mL) was then added dropwise, and the resulting mixture stirred at room temperature for 2 hours, after which it was poured into water, extracted and washed as described above and dried. After filtration and removal of solvent, the residue was purified by column chromatography over silica gel, eluting with 50% ethyl acetate in methylene chloride. The product was obtained as red liquid ( 4.5g, 63% ); 1H NMR δ: 1.55 (s, 1 H, OH), 1.9 (m, 2 H, CH₂-CH₂-OH), 3.03 (t, 2 H, J=7.1 Hz, S-CH₂), 3.76 (t, 2 H, J=6 Hz, CH₂ -OH), 6.37 (dd, 1 H, J=15.6 Hz, J=7.7 Hz, =CHCHO), _2 7.01 (d, 1 H, J=3.8 Hz, aromatic H), 7.18 (d, 1 H, J=3.8 Hz, aromatic H), 7.44 (d, 1 H, J=15.6 Hz, vinyl), 9.57 (d, 1 H, J=7.7 Hz, CHO); ^_ax /nm (ε_max/dm³ mol^"1 cm^"1) 354.5 (17 400) HRMS (El) Calc. for CιoHι₂O₂S₂288.0281, found 288.0279. The product was used without further purification in the preparation of 65.

EXAMPLE 11 - Preparation of Thienyl-Containing Chromophores

l-[5'-Butylthio-2'-thienyl]-2-[5"-(3'"-hydroxypropylthio)-2"-thienyl] ethene (64)

A solution of sodium ethoxide (1 M in ethanol, 0.062mol, 62 mL ) was added dropwise to a solution of 62 (6g, 0.025 mol ) and 48 (13.78g, 0.03 mol ) in ethanol (75 mL ) at room temperature. The resulting mixture was stirred at 60 °C overnight, after which it was cooled to room temperature. After removal of ethanol, the residue was purified by column chromatography over silica gel, eluting with methylene chloride. The product was obtained as yellow solid (8.7g, 96% ), m.p. 56-57 °C, 1H NMR δ: 0.89 (t, 3 J-7.3 Hz, CH3), 1.36 (s, 1 H, OH), 1.39 (m, 2 H, CH2-CH3), 1.59 (m, 2 H, CH2-CH2-CH3),1.88 (m, 2 H, CH2- CH2-OH), 2.8 (t, 2 H, J=7.3 Hz, S-CH2), 2.92 (t, 2 H, J=7.1 Hz, S-CH2- CH2-CH2-OH), 3.76 (t, 2 H, J=5.9 Hz, CH2-OH), 6.46-6.97 (m, 6 H, vinyl and aromatic H); ¹³C NMR: 14.05, 22.03, 31.92, 32.44, 35.6, 38.81, 61.55, 121.67, 122.05, 122.73, 123.37, 126.97, 127.09, 133.74, 134.31, 145.01, 145.34; _λmax /nm (_εmaχ/dm³ mol^"1 cm^"1) 368 (18 700); (Anal.Calc. for d₇H₂₂OS₄: C, 55.09; 11, 5.99. Found: C, 55.09; H, 6.02)

l-[5'-Butylthio-2'-thienyl]-4-[5"-(3"'-hydroxypropylthio)-2"-thienyl]-l,3- butadiene (65) A solution of sodium ethoxide (1 M in ethanol, 0.024 mol, 24 mL ) was added dropwise to a solution of 63 (2.7g, 0.012 mol ) and 48 (6.73g, 0.014 mol ) in ethanol (75 mL) at room temperature. The resulting mixture was stirred at 60 °C overnight after which it was cooled to room temperature. After removal of ethanol, the residue was purified by column chromatography over silica gel, eluting with methylene chloride. The product was obtained as yellow solid ( 4 g, 85% ), m.p. 72.5-74.5 °C, 1H NMR δ: 0. 89 (t, 3H, J=7.3 Hz, CH3), 1.41 (m, 2 H, CH3-CH2), 1.6 (m, 2 H, CH3-CH2-CH2), 1.87 (m, 2 H, CH2-CH2-OH), 2.8 (t, 2 H, J=7.3 Hz, CH2-S), 2.91 (t, 2 H, J=7.1 Hz, S-CH2), 3.76 (t, 2 H, J=6.1 Hz, CH2-OH), 6.57-6.96 (m, 8 H, aromatic and vinyl H); ¹³C NMR: 14.05, 22.03, 31.92, 32.44, 35.60, 38.81, 61.55, 125.77, 126.08, 126.82, 126.90, 128.90, 129.23, 133.78, 134.21, 134.34, 135.22, 146.00, 146.50 ; _λmax /nm G_maχ/dm³ mol^"1 cm^"1) 390 (45 000); (Anal.Calc. for Cι₉H₂₄OS₄: C, 57.53; H, 6.10. Found: C, 57.41; H, 6.12)

l-[5'-Butylthio-2'-thienyl]-2-[5"-(3'"-iodopropylthio)-2"-thienyl] ethene (66)

Iodine ( 6.17g, 0.025 mol ) was added slowly to a solution of triphenylphosphine (6.38g, 0.025 mol) and imidazole ( 1.65g, 0.025 mol ) in a 1 :3 mixture of acetonitrile: ether ( 100 mL ) at 0 °C. The ice bath was removed and the mixture was stirred for 15 minutes. Compound 64 (3g, 0.008 mol ) in 1 :3 acetonitrile: ether mixture (20 mL) was added dropwise. The resulting mixture was stirred for an hour at room temperature. After removal ofthe solvent the residue was purified by column chromatography over silica gel, eluting with 5% ethyl acetate in hexanes to yield 66 { 3.7g, 95.8% ) The product was obtained as gel, therefore melting point could not be determined. H NMR δ: 0. 89 (t, 3 H, J=7.3 Hz, CH₃), 1.41 (m, 2 H, CH2-CH2), 1.6 (m, 2 H, CH2- CH2-CH3), 2.08 (m, 2 H, CH2-CH2-I), 2.81 (t, 2 H, J=7.3 Hz, S-CH2), 2.88 (t, 2 H, J=6.9 Hz, S-CH2), 3.28 (t, 2 H, J=6.7 Hz, Clfc-I), 6.84-6.97 (m, 6 H, aromatic and vinyl H); _λmaχ/nm (_εmaχ/dm³ mol^"1 cm^"1) 373 (45 500); (Anal.Calc. for Cι₇H₂ιIS₄: C, 42.45; H, 4.51. Found: C, 42.70; H, 4.34) The product was used without further purification in the preparation of 69.

l-[5'-Butylthio-2^,-thienyl]-4-[5"-(3'"-iodopropylthio)-2"-thienyl]-l,3- butadiene (67)

Compound 67 was prepared from 65 ( 2g, 0.005 mol ), iodine ( 3.85g, 0.0 15 mol), triphenyl phosphine ( 3.94g, 0.015 mol ) and imidazole ( 1.02g, 0.0.15 mol ) as described above for the preparation of 66. The crude product was purified by column chromatography over silica gel, eluting with 5% ethyl acetate in hexanes to yield 67 ( 2.4g, 95% ) The product was obtained as gel, therefore the melting point could not be determined. ^!H NMR δ: 0.89 (t, 3 H, J=7.3 Hz, CH3),1.4 (m, 2 H, CH2-CH3),1.59 (m, 2 H, CH2-CH2-CH3), 2.09 (m, 2 H, CH2-CH2-I), 2.8 (t, 2 H, J=7.3 Hz, S-CH2), 2.87 (t, 2 H, J=6.9 Hz, S- CH2), 3.28 (t, 2 H, J=6.7 Hz, CH2I), 6.54-6.96 (m, 8 H, aromatic and vinyl H); _λmax/nm (_εma_x/dm³ mol^"1 cm^"1) 390 (66 600); (Anal.Calc. for Cι₉H₂₃IS₄: C, 45.05; H, 4.58. Found: C, 45.70; H, 4.58) The product was used without further purification in the preparation of 69.

3,5-Bis-[2' (5"-butylthio-2"— thienyl)-r-(5"'-thiopropyleneoxy-2^,π- thienyl)ethene] benzyl alcohol (68) A mixture of 66 ( 3.72 g, 7.74 mmol), 3,5-dihydroxybenzyl alcohol

(Aldrich) (0.53 g, 3.78 mmol ), potassium carbonate (1.05g, 7.56 mmol ) and 18-Crown-6 (Aldrich) ( 0.2 g, 0.78 mmol) in dry 1,4-dioxane ( 75 mL ) was refluxed under a nitrogen atmosphere for 48 hours. The mixture was cooled and evaporated to dryness. The residue was proportioned between methylene chloride (50 ml,) and water (50 mL). The aqueous layer was extracted with methylene chloride (3 X 75 mL ). The combined organic layers were dried over MgSO . After filtration and removal of solvent, the residue was purified by column chromatography over silica gel, eluting with methylene chloride to yield 68. The product was obtained as gel, therefore melting point could not be determined. ( 2.07 g, 65%) 1H NMR δ: 0.89 (t, 6 H, J=7.3 Hz, CH3), 1.41 (m, 4 H, CH2-CH3), 1.55 (s, 1 H, OH), 1.6 (m, 4 H, CH2-CH2-CH3), 2.06 (m, 4 H, CH2- CH2-O), 2.8 (t, 4 H, J=7.3 Hz, S-CH2), 2.97 (t, 4 H, J-7 Hz, S-CH2), 4.03 (t, 4 H, J=5.9 Hz, CH2O), 4.59 (d, 2 H, J=5.7 Hz, CH2-OH), 6.33 (s, 1 H, phenyl H), 6.48 (s, 2 H, phenyl H), 6.82-6.96 (m, 12 H, aromatic and vinyl H); ¹³C NMR: 14.05, 22.03, 29.47, 31.92, 35.57, 38.80, 65.71, 66.21, 101.04, 105.67, 121.66, 122.07, 122.72, 123.25, 126.97, 127.12, 133,73, 134.42,

143.79, 145.32, 145.97, 160.57; λ™_* /nm (ε_max/dm³ mol^"1 cm^"1) 373 (54 700); (Anal.Calc. for C₄ιH₄₈O₃S₈: C, 58.25; H, 5.72. Found: C, 58.39; H, 5.71)

3,5-Bis-[4'-(5"-butylthio-2"-thienyl)-l'-(5"^,-thiopropyleneoxy-2^,"-thienyl)- 1,3-butadiene] benzyl alcohol {69)

A mixture of 67 ( 1.9 g, 3.75 mmol ), 3,5-dihydroxybenzyl alcohol (0.25 g, 1.79 mmol ), potassium carbonate ( 0.5 g, 3.58 mmol ) and 18-Crown-6 ( 0.1 g, 0.36 mmol ) in dry 1,4-dioxane ( 75 mL ) was refluxed under a nitrogen atmosphere for 48 hours. The mixture was cooled and evaporated to dryness. The residue was proportioned between methylene chloride (50 mL) and water (50 mL). The aqueous layer was extracted with methylene chloride (3 X 75 mL ). The combined organic layers were dried (MgSO₄) and after filtration and removal of solvent, the residue was purified by column chromatography over silica gel, eluting with ethylene chloride to yield 69. The product was obtained as gel, therefore, melting point could not be determined. (1. 15 g, 76%); 1H NMR δ: 0. 92 (t, J=7.2 Hz, CH3), 1.43 (m, 4 H, CH2-CH3), 1.61 (m, 4 H, CH2-CH2CH3), 2.09 (m, 4 H, CH2-CH2-O), 2.82 (t, 4 H, J=7.3 Hz, S-CH2), 2.99 (t, 4 H, J=7 Hz, S-CH2), 4.06 (t, 4 H, J=5.8 Hz, CH2-O), 4.62 (s, 2 H, CH2-O), 6.36 (s, 1 H, phenyl H), 6.5 (s, 2 H, phenyl H), 6.55-6.98 (m, 16 H, aromatic and vinyl H); /nm (ε_max/dm³ mol^"1 cm^"1) 390 (86 800); ¹³C NMR: 14.02, 22.02, 29.45, 31.92, 35.56, 38.80, 65.76, 66.22, 101.05, 105.68, 125.74, 126.11, 126.80, 126.91, 128.87, 129.25, 133.78, 134.11, 134.45, 135.23, 143.75, 145.98, 146.59, 160.59 ; (Anal.Calc. for C₄₅H₅₂O₃S₈: C, 60.23; H, 5.84. Found: C, 60.28; H, 5.88)

EXAMPLE 12 - Preparation of Triphenylamine Derivatives

4-(N,N-diphenylamino)benzyl acetate

4-(N,N-Diphenylamino)benzyl alcohol (5.94g, 0.0216 mol), pyridine

(16 mL) and acetic anhydride (11.4 mL) were mixed in a three-neck flask equipped with a condenser and addition funnel. The resulting mixture was stirred overnight at room temperature. Deionized water (50 mL) was then added and the resulting mixture stirred for 2 hours. The product was obtained by filtration, and recrystallized from hexane to yield the desired product (6.52g, 95%), m. p. 105-6 °C, lH NMR δ 2. 10 (s, 3H, CH3), 5.04 (s, 2H, CH2), 7.0-7.4 (m, 14H, phenyl). Analysis: Calculated for Cι₉Hι₇NO₂,: C, 79.46%, H, 6.03%, N, 4.41%; Found: C, 79.37 %, H, 5.72 %, N, 4.41%.

4- [4 '-Formyl-N.N-diphenylamino] b enzyl acetate

DMF (2.63g, 0.036 mol) was added to a three-neck flask (50 mL) equipped with a condenser, CaCl₂ drying tube, and a glass stopper and cooled in an ice bath. Phosphorus oxychloride (1.8g, 0.012 mol) was then added dropwise, and the resulting mixture stirred for 5 minutes and then allowed to warm slowly to room temperature. A solution of 4-(N,N-diphenylamino)benzyl acetate (1.88g, 0.0059 mol) in DMF (20 mL) was then added dropwise. After addition was complete, the mixture was heated to 70°C and stirred overnight. The mixture was then poured over ice (20 g) and the product obtained by filtration after slowly neutralizing the solution in an ice bath with saturated sodium acetate solution to pH 6. After washing with deionized water and drying, the product was recrystallized by dissolving in a small quantity of chloroform and pouring into excess hexane to yield the desired product (1.21g, 60%), m. p. 86-89°C, 1H NMR δ 2.12(s, 2H, CH3), 5,09 (s, CH2), 7.0-7.37 (m, 12H, phenyl), 7.65-7.70 (d, 2H, phenyl), 9.81 (s, 1H, CHO). Analysis:

Calculated for C₂₂Hι₉NO₃: C, 76.52%, H, 5.51%, N, 4.06%; Found: C, 76.27%, H, 5.72%, N, 4.01%.

3-[4'-(4"-Acetoxymethyl-N,N-diphenylamino)phenyl]-2-propenal

A solution of NaOEt in ethyl alcohol (1M, 0.03 mol, 30 mL) was added dropwise to a solution of 4-[4'-Formyl-N.N-diphenylamino]benzyl acetate (3.45g, 0.01 mol) and tributyl(l,3-dioxolan-2-ylmethyl)phosphonium bromide (0.54 M in DMF, 0.03 mol, 30 mL) in DMF (20 mL). The temperature ofthe mixture was raised to 90 °C and stirred overnight. After cooling the solution to room temperature, aqueous HCI (3M, 30 mL) was added dropwise, the mixture stirred for 1 hour, poured into ice (75 g), and the product extracted with ethyl ether (3 x 30 mL). The combined extracts were washed with saturated Na₂CO₃ solution and brine, dried over MgSO₄ and the solvent evaporated. The product was purified by dissolving in warm ethanol, filtering, and evaporating the solvent to yield the desired product (2.80g, 81%) m. p. 112-115°C, 1H NMR δ 2.0 (s, 3H, CH3), 4.68 (s, 2H, CH2O),6.53-6.64 (dd, 2H, vinyl), 7.0-7.4 (m, 13H, phenyl), 9.62-9.65 (d, 1H, CHO). Analysis: Calculated for C₂₄H₂ιNO₃: C, 77.49%, H, 5.82%, N, 3.76%; Found: C, 77.38%, H, 5.54%, N, 3.62%.

4-[(4',4"-Dibromo)diphenylamino]benzaldehyde

Triphenylamine-4-carbaldehyde (22. Og, 0.08 mol) was dissolved in chloroform (100 mL) in a IL Erlenmeyer flask. Glacial acetic acid (250 mL) was then added and the resulting bilayer was stirred while bromine (8.66 mL, 0. 169 mol, 2.1 equiv.) was added dropwise via syringe while maintaining the reaction temperature at 25 °C. After the addition was complete, the reaction mixture was stirred for 1 hour, after which water (250 mL) was added to the reaction mixture. The organic and aqueous layers were separated, and the aqueous layer was extracted with dichloromethane (2 x 75 mL). The combined organic product solution was then washed with an aqueous solution of KOH (1 M, 100 mL), followed by water (4 x 100 mL) and brine (100 mL), and was then dried with MgSO₄. The solvent was removed by rotovaporization and the resulting green solid was recrystallized from acetonitrile to afford yellow crystals (33.4g, 96%), IH NMR δ 7.75-7.22 (m, 12 H, Ar-H), 9.81 (s, IH, CHO).

4-(Diphenylamino)benzyl alcohol

Triphenylamine-4-carbaldehyde (93g, 0.34 mole) dissolved in anhydrous THF (1 equiv.) was added dropwise to a suspension of LiAlH₄ (0.34 mol) in anhydrous THF contained in a 2L 3-neck flask equipped with an addition funnel, magnetic stir bar, reflux condenser and stopper maintained at 0 °C. After the addition was complete, the reaction mixture was allowed to warm to 25 °C, and was stirred for 2 hours. The reaction progress was monitored by TLC. After the reaction was complete, the mixture was again cooled to 0 °C, and water (200 mL) was added dropwise via the addition funnel to consume any residual LAH. An aqueous solution of HCI (2M, 200 mL) was then added, and the resulting emulsion was stirred for 10 minutes, and then extracted with dichloromethane (3 x 150 mL). The combined organic product was then washed with water (100 mL) followed by brine (100 mL) and dried with MgSO₄. The solvent was removed by rotovaporization to yield the crude product. The product was recrystallized from hexane to yield, the desired product, m. p. 104-5 °C, IH NMR δ 4.64 (s, 2H, CH2OH), 7.01-7.27 (m, 14 H, Ar-H); 13C NMR 123.2, 124.4, 124.6, 125.0, 128.7, 129.6, 135.4, 148.1 ppm. Diethyl (4-diphenylamino)benzyl phosphonate

A IL round bottom flask equipped with a magnetic stir bar was charged with a solution of triphenylphosphine (7.2g, 0.0275 mol, 1. 1 eq) in ethyl ether. Bromine (1.4 mL, 0.0275 mol, 1 eq) was then added via syringe to the stirred solution. A saturated solution of 4(diphenylamino)benzyl alcohol (6.96g, 0.025 mol, 1 eq) in ethyl ether was poured into the reaction mixture and stirred for 1.5 hoxir. A white precipitate formed and was isolated by vacuum filtration. The filtered organic layer was then rotovaporized to yield a red-brown gum, which was then dissolved in toluene (250 mL). Triethylphosphite (4.28 mL, 0.025 mol, 1 eq) was added to the toluene solution, and the resulting mixture was placed in a 500 mL round bottom flask equipped with a reflux condenser, and heated at 120 °C for 16 hours. The reaction was monitored by TLC (4: 1 dichloromethane: ethyl acetate). After completion ofthe reaction, the solvent was removed by rotovaporization and the light yellow liquid product was further purified by column chromatography (4:1 dichloromethane: ethyl acetate) to afford the desired product as a white solid (9.2g, 93%), IH NMR δ 1.24-1.34- (t, 6H, J 14 Hz, CH3CH2OP), 3.04-3.11 (dq, 4H, Jab 15 Hz, Jbc 14.6 Hz,

CH3CH2OP), 3.97-4.11 (d, 2H, J 21 Hz, ArCH2OP), 6.95-7.21(m, 14H, Ar-H).

4-[(4',4"-Dibromo)diphenylamino]benzyl alcohol

Reduction of 4-[(4',4"-dibromo)diphenylamino]benzaldehyde (0.0435 mol) with LAH (0.0435 mol) was carried out as described above for 4-(diphenylamino)benzyl alcohol, to give crude

4-[(4',4"-dibromo)diphenylamino]benzyl alcohol which was purified by silica gel column chromatography eluting with dichloromethane/ethyl acetate (4: 1) to give pure 4-[(4',4"-dibromo)diphenylamino]benzyl alcohol (34.4g, 91 %), IH NMR δ 4.62 (s, 2H, CH2OH), 6.77-7.40 (m, 12H, Ar-H).

4-(4'hydroxymethyl)diphenylaminobenzaldehyde and 4-(hydroxymethyl)-4',4"diformyItriphenylamine

A 3-neck round bottom flask equipped with a nitrogen inlet, drying tube, magnetic stir bar and addition funnel was charged with a solution of 4-[(4',4"-Dibromo)diphenylamino]benzaldehyde (25.08g, 0.0577 mol, 1 eq.) in anhydrous THF (600 mL). The solution was cooled to -100 °C. n-Butyllithium in hexane (2.5M, 115 mL, 0.289 mol, 5 eq) was transferred to the addition funnel via cannula, and then added dropwise. The temperature was then allowed to rise slowly to -80 °C, and DMF (27 mL, 0.346 mol, 6 eq.) in a clean addition funnel was then added dropwise to the reaction mixture. The cooling bath was then removed and the reaction mixture was allowed to warm slowly to room temperature. The progress ofthe reaction was monitored by TLC (9:1 dichloromethane: ethyl acetate). After completion, an aqueous solution of HCI (2M, 100 mL) was added to the solution, and the aqueous layer was separated from the organic layer, and further extracted with ethyl ether (100 mL). The combined organic product was washed with water (3 x 100 mL) and once with brine, and then dried with MgSO₄. After removal ofthe solvent by rotovaporization, the crude mixture was purified by silica gel column chromatography (9:1 dichloromethane: ethyl acetate) yielding two pure products, 4-(4'hydroxymethyl)diphenylaminobenzaldehyde (6.17g, 35%) and 4-(hydroxymethyl)-4',4"diformyltriphenylamine (7.34g, 37 %), 4-(4'hydroxymethyl)diphenylaminobenzaldehyde: IH NMR δ 4.65 (s, 2H, CH2OH), 6.87-7.79 (m, 13H, Ar-H), 9.87 (s, IH, CHO); and 4-(hydroxymethyl)-4',4"diformyltriphenylamine: IH NMR δ 4.66 (s, 2H, CH2OH), 7.1-7.81 (m, 12H, Ar-H), 9.87 (s, 2H, CHO). Although the foregoing description is directed to the preferred embodiments ofthe invention, it is noted that other variations and modifications will be apparent to those skilled in the art, and may be made without departing from the spirit or scope ofthe invention. The complete disclosures of all references cited above are hereby incoφorated by reference in their entireties for all pmposes.

Claims

WHAT WE CLAIM IS:

1. A poφhyrin-based compound comprising the following poφhyrin stracture:

wherein each R¹ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, - CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; each R² is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂,

-(CH₂O)n-G, -t-Butyl, or -C(O)OG; each R³ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; or independently R² and R³ are linked by • C₄H₄- to form a six-member ring; M is either two hydrogen atoms or a metal ion; each n is independently an integer ranging from 1 to 20; each G is independently -H, or a d to C₂₀ alkyl;

Ar comprises one of C₆H₅, C₆H X, C₆H₃X₂, C₆H₂X₃, CβFD , or C₆X₅, wherein X comprises one of F, Cl, Br, or I; L is a linking moiety between the poφhyrin structure and TPA; TPA is a chromophore moiety; and wherein at least one of R¹, R² or R³ is a L-TPA moiety, and at least one other of R¹, R² or R³ is not H or a L-TPA moiety.

2. A compound according to claim 1, wherein the TPA moiety is attached to the poφhyrin ring directly or with a linking group, L.

3. A compound according to claim 2, where the linking group comprises one linking moiety selected from the group consisting of ethenyl, ethynyl, -

(CH₂)n- wherein n is equal to 1 to 20, ortho-phenyl, meta-phenyl, para-phenyl, - C(=O)-O-, and 4-phenyl-2' -ethenyl.

4. A compound according to claim 1, wherein the TPA moiety comprises one member selected from the group consisting ofthe following structures:

TPA A

TPA B

wherein in TPA B, n = 1 to 5, and R comprises an alkyl;

TPA C

TPA E wherein in TPA E, n = 1 to 5, R comprises one member selected from the group consisting of H, alkyl, and -(OCH CH₂)_nOG; wherein G is H or alkyl, and R' comprises alkyl;

TPA F

TPA G

TPA H

wherein in TPA H, R comprises one member selected from the group consisting of alkyl, phenyl, alkyloxyphenyl, phenyl[(OCH₂CH₂)nOG]x; wherein G is H or alkyl, n=l to 4, and x=l to 4; wherein alkyl comprises C_\ to C₂₀ alkyl moieties; wherein the selected TPA is attached to the poφhyrin structure at the point indicated by the wiggle line.

5. A compound according to claim 1, wherein the compound comprises 5- [4"'-(diphenylamino)-4"-stilbenyl]-15-[2',6'-dichlorophenyl]-21H, 23H- poφhyrin.

6. A compound according to claim 1, wherein M comprises a metal atom.

7. A compound according to claim 1, wherein M comprises two hydrogen atoms.

8. A photodynamic therapy agent comprising a compound according to any one of claims 1 to 7.

9. A method of increasing the multi-photon absoφtion cross-section of a poφhyrin-based photosensitizer comprising attaching at least one TPA-chromophore at the meso or beta positions of a poφhyrin stracture ofthe poφhyrin-based photosensitizer, and attaching at least one intersystem crossing enhancing substituent to meso or beta positions of a poφhyrin structure ofthe poφhyrin-based photosensitizer, whereby the increased multi-photon absoφtion cross-section ofthe poφhyrin-based photosensitizer is at least about 30 GM units at about its maximum wavelength for two-photon absoφtion.

10. A method according to claim 9, wherein the TPA-chromophore comprises one member selected from the group consisting ofthe following structures:

TPA A wherein in TPA A, n = 1 to 5, and R comprises one member selected from the group consisting of H, alkyl, alkyloxy, -(OCH₂CH₂)nOG; wherein G is H or alkyl;

TPA B

wherein in TPA B, n = 1 to 5, and R comprises an alkyl;

TPA C wherein in TPA C, n = 1 to 3, R comprises one member selected from the group consisting of H, CN, alkyl, alkyloxy, and R' comprises one member selected from the group consisting of alkyl, alkyloxyphenyl, phenyl, phenyl- (OCH₂CH₂)nOG; wherein G is H or alkyl;

TPA D

TPA E wherein in TPA E, n = 1 to 5, R comprises one member selected from the group consisting of H, alkyl, and -(OCH CH )_nOG; wherein G is H or alkyl, and R' comprises alkyl;

TPA F

TPA G

TPA H

wherein in TPA H, R comprises one member selected from the group consisting of alkyl, phenyl, alkyloxyphenyl, phenyl[(OCH₂CH₂)nOG]x ; wherein G is H or alkyl, n=l to 4, and x=l to 4; wherein in TPA's A to H, alkyl comprises Ci to C ₀ alkyl moieties; and wherein the TPA is attached to the poφhyrin ring at the point indicated by the wiggle line.

11. A method according to claim 10, wherein the TPA-chromophore is attached to the poφhyrin ring directly or with a linking group.

12. A method according to claim 11, where the linking group comprises one linking moiety selected from the group consisting of ethenyl, ethynyl, -(CH2)n- wherein n is equal to 1 to 20, ortho-phenyl, meta-phenyl, para-phenyl, -C(=O)- O-, and 4-phenyl-2' -ethenyl.

13. A method according to claim 9, wherein the at least one intersystem crossing enhancing substituent comprises one member selected from the group consisting of C₆H₅, CβBUX, C₆H₃X₂, C₆H X₃, C₆HX₄, or C₆X₅, wherem X comprises one of F, Cl, Br, or I.

14. A method according to claim 9, wherein the increased multi-photon absoφtion cross-section ofthe poφhyrin-based photosensitizer is at least about 50 GM units at about its maximum wavelength for two-photon absoφtion.

15. A method according to claim 9, wherein the increased multi-photon absoφtion cross-section ofthe poφhyrin-based photosensitizer is at least about 70 GM units at about its maximum wavelength for two-photon absoφtion.

16. A method according to claim 9, wherein the poφhyrin structure of the poφhyrin-based photosensitizer comprises at least one member selected from the group consisting of poφhyrin, chlorin, bacteriochlorin, and isobacteriochlorin.

17. A method according to claim 9, wherein the poφhyrin-based photosensitizer absorbs two photons of radiation in the range of about 700 nm to about 1300 nm.

18. A method according to claim 17, wherein the absorbed radiation is in a range of about 700 nm to about 1100 nm.

19. In an improved method of conducting photodynamic therapy which comprises providing a therapeutically effective amount of a photoactive compound to within a subject, contacting the photoactive compound with an area to be treated within the subject, and exposing the area to radiation with a wavelength ranging from between about 700 nm to about 1300, wherein the improvement comprises a providing a therapeutically effective amount ofthe photoactive compound the following poφhyrin structure:

wherein each R¹ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, - CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; each R² is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂, -

(CH₂O)n-G, -t-Butyl, or -C(O)OG; eachR³ is independently -L-TPA, -H, -Ar, -(CH₂)nCH₃, -CH=CH₂, -(CH₂O)n-G, -t-Butyl, or -C(O)OG; or independently R² and R³ are linked by -C₄H₄- to form a six-member ring; M is either two hydrogen atoms or a metal ion; each n is independently an integer ranging from 1 to 20; each G is independently -H, or a Cl to C20 alkyl;

Ar comprises one of C₆H₅, C₆H X, C₆H₃X₂, C₆H₂X , C₆HX₄, or C₆X₅, wherein X comprises one of F, Cl, Br, or I; L is a linking moiety between the poφhyrin structure and TPA;

TPA is a chromophore moiety; and wherein at least one of R¹, R² or R³ is a L-TPA moiety, and at least one other of R¹, R² or R³ is not H or a L-TPA moiety.

20. A method according to claim 19, wherein the TPA moiety is attached to the poφhyrin ring directly or with a linking group, L.

21. A method according to claim 20, where the linking group comprises one linking moiety selected from the group consisting of ethenyl, ethynyl, -(CH₂)n- wherein n is equal to 1 to 20, ortho-phenyl, meta-phenyl, para-phenyl, -C(=O)- O-, and 4-phenyl-2' -ethenyl.

22. A method according to claim 19, wherein the TPA moiety comprises one member selected from the group consisting ofthe following structures:

TPA A

TPA B

wherein in TPA B, n = 1 to 5, and R comprises an alkyl;

TPA C

TPA F

TPA G

TPA H

wherein in TPA H, R comprises one member selected from the group consisting of alkyl, phenyl, alkyloxyphenyl, phenyl[(OCH₂CH₂)nOG]x; wherein G is H or alkyl, n=l to 4, and x=l to 4; wherein alkyl comprises d to C₂₀ alkyl moieties; wherein the selected TPA is attached to the poφhyrin structure at the point indicated by the wiggle line.

23. A method according to claim 19, wherein the compound comprises 5- [4"'-(diphenylamino)-4"-stilbenyl]-15-[2',6^,-dichlorophenyl]-21H, 23H- poφhyrin. '

24. A method according to claim 19, wherein M comprises a metal atom.

25. A method according to claim 19, wherein M comprises two hydrogen atoms.

26. A method according to claim 19, wherein a therapeutically effective amount comprises an amount ranging from about 0.1 mg to about 10 mg per kg of body weight.