Patent Application
20210309645
The present application relates to photoacoustic compounds and, in particular, to photoacoustic compounds having pH dependent absorption spectra.
While near infrared (NIR) fluorescent intraoperative imaging systems are able to provide simultaneous acquisition of surgical anatomy and NIR fluorescence signal, two major impediments, limited depth of resolution (5-8 mm) below the surface and limited cancer-specific probes with sufficient signal, restrict the potential for image-guided surgical resection to more superficial tumors, i.e. melanoma, some superficial head and neck squamous cell carcinoma, other oral cancers, and some thyroid or breast cancers. While intraoperative imaging strategies to visualize cancer cells accurately are necessary, it is critical to develop intraoperative imaging methods that can detect molecular information at depths of centimeters for non-superficial cancers, while retaining high resolution and tumor specificity.
Photoacoustic (optoacoustic) imaging is emerging to drive optical imaging beyond the penetration limits of conventional methods by allowing the formation of optical images several centimeters inside tissue. Multispectral optoacoustic imaging is based on the optoacoustic effect: the conversion of absorbed electromagnetic energy (e.g. NIR light) to acoustic signals. The selective absorption of light at multiple wavelengths and slices enables 3D volumetric spectrally enriched (color) imaging from deep living tissues in real time and at high spatial resolution. MSOT imaging operates through centimeters of tissue enabling tomographic 3D imaging with optical contrast at depths of ultrasound, in real-time. Currently, clinical Multispectral Optoacoustic imaging systems are in testing to improve tumor identification in Europe and recently the United States. To maximize the potential of clinical MSOT imaging, tumor specific contrast agents must be developed.
In one aspect, photoacoustic compounds or optoacoustic compounds are described herein. Photoacoustic and optoacoustic are generally used interchangeably in the art. A photoacoustic compound, for example, comprises two multicyclic ring moieties coupled by a rotationally restricted linkage, wherein the photoacoustic compound exhibits at least one of a pH dependent absorption spectrum, pH dependent emission spectrum and pH dependent photoacoustic spectrum. In being pH dependent, the absorption, emission and/or photoacoustic spectra of the photoacoustic compound can vary in response to environmental pH changes. Accordingly, the photoacoustic compound can provide differing photoacoustic responses based on the local pH environment. In this way, various targets can be imaged and resolved based on pH variance between the targets.
In some embodiments, a photoacoustic compound described herein is of Formula (I):
wherein R1-R19 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, hydroxyl, halo, and amide, wherein the alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, and amide are optionally substituted with one or more substituents selected from the group consisting of (C1-C10)-alkyl, (C1-C10)-alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amide, halo, hydroxy, C(O)OR20, and C(O)R21, wherein R20 is selected from the group consisting of hydrogen, alkyl and alkenyl and R21 is selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl and NR22R23, wherein Rn and R23 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and heteroaryl; and wherein X is selected from the group consisting of CR24R25, N and O, wherein R24 and R25 are independent selected from the group consisting of hydrogen, alkyl, alkenyl, hydroxyl and halo.
In some embodiments, a photoacoustic compound described herein is of Formula (II):
wherein R1-R17 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, hydroxyl, halo, and amide, wherein the alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, and amide are optionally substituted with one or more substituents selected from the group consisting of (C1-C10)-alkyl, (C1-C10)-alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amide, halo, hydroxy, C(O)OR18, and C(O)R19, wherein Rig is selected from the group consisting of hydrogen, alkyl and alkenyl and Rig is selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl and NR20R21, wherein R20 and R21 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and heteroaryl; and wherein X is selected from the group consisting of CR22R23, N and O, wherein R24 and R25 are independent selected from the group consisting of hydrogen, alkyl, alkenyl, hydroxyl and halo.
In another aspect, methods of imaging are described. Briefly, a method of imaging a target comprises disposing a photoacoustic compound in an environment adjacent to the target and irradiating the photoacoustic compound with one or more wavelengths of electromagnetic radiation. Soundwaves produced by the photoacoustic compound in response to the electromagnetic radiation are detected and transformed into an image. The photoacoustic compound comprises two mutlicyclic ring moieties coupled by a rotationally restricted linkage, and exhibits at least one of a pH dependent absorption spectrum, pH dependent emission spectrum and pH dependent photoacoustic spectrum. In some embodiments, the photoacoustic compound is irradiated with multiple wavelengths of radiation resulting in multiple wavelength dependent images of the target. As described further herein, each of the wavelength dependent images can correspond to the photoacoustic compound at locations of differing pH in the environment. The wavelength dependent images can be overlaid to present a detailed image of the target.
These and other embodiments are described in greater detail in the following detailed description.
Embodiments described herein can be understood more readily by reference to the following detailed description and examples and their previous and following descriptions. Elements, apparatus and methods described herein, however, are not limited to the specific embodiments presented in the detailed description and examples. It should be recognized that these embodiments are merely illustrative of the principles of the present invention. Numerous modifications and adaptations will be readily apparent to those of skill in the art without departing from the spirit and scope of the invention.
The term “alkyl” as used herein, alone or in combination, refers to a straight or branched saturated hydrocarbon group optionally substituted with one or more substituents. For example, an alkyl can be C1-C30 or C1-C18.
The term “alkenyl” as used herein, alone or in combination, refers to a straight or branched chain hydrocarbon group having at least one carbon-carbon double bond and optionally substituted with one or more substituents.
The term “aryl” as used herein, alone or in combination, refers to an aromatic monocyclic or multicyclic ring system optionally substituted with one or more ring substituents.
The term “heteroaryl” as used herein, alone or in combination, refers to an aromatic monocyclic or multicyclic ring system in which one or more of the ring atoms is an element other than carbon, such as nitrogen, oxygen and/or sulfur.
The term “cycloalkyl” as used herein, alone or in combination, refers to a non-aromatic, mono- or multicyclic ring system optionally substituted with one or more ring substituents.
The term “heterocycloalkyl” as used herein, alone or in combination, refers to a non-aromatic, mono- or multicyclic ring system in which one or more of the atoms in the ring system is an element other than carbon, such as nitrogen, oxygen or sulfur, alone or in combination, and wherein the ring system is optionally substituted with one or more ring substituents.
The term “heteroalkyl” as used herein, alone or in combination, refers to an alkyl moiety as defined above, having one or more carbon atoms in the chain, for example one, two or three carbon atoms, replaced with one or more heteroatoms, which may be the same or different, where the point of attachment to the remainder of the molecule is through a carbon atom of the heteroalkyl radical.
The term “alkoxy” as used herein, alone or in combination, refers to the moiety RO—, where R is alkyl or alkenyl defined above.
The term “halo” as used herein, alone or in combination, refers to elements of Group VIIA of the Periodic Table (halogens). Depending on chemical environment, halo can be in a neutral or anionic state.
In one aspect, photoacoustic compounds are described herein. A photoacoustic compound, for example, comprises two multicyclic ring moieties coupled by a rotationally restricted linkage, wherein the photoacoustic compound exhibits at least one of a pH dependent absorption spectrum, pH dependent emission spectrum and pH dependent photoacoustic spectrum. In being pH dependent, the absorption, emission and/or photoacoustic spectra of the photoacoustic compound can vary in response to environmental pH changes. Accordingly, the photoacoustic compound can provide differing photoacoustic responses based on the local pH environment.
As detailed further herein, photoacoustic compounds can be zwitterionic. In some embodiments, the two multicyclic ring moieties are oppositely charged. Alternatively, one of the two multicyclic ring moieties can possess both positive and negatively charged groups. Moreover, each of the multicyclic ring moieties can exhibit conjugation and/or aromaticity. In some embodiments, at least one of the multicyclic ring moieties is aromatic. In such embodiments, the other multicyclic moiety coupled to the rotationally restricted linkage is also aromatic or partially conjugated. The rotationally restricted linkage between the two multicylic moieties can render the photoacoustic compound planar or substantially planar. However, the photoacoustic compound can exhibit out of plane flexing. In some embodiments, the photoacoustic compound exhibits out of plane flexing in response to exposure to electromagnetic radiation of one or more wavelengths. The rotationally restricted linkage between the multicyclic moieties can comprise an unsaturated hydrocarbon, in some embodiments. The unsaturated hydrocarbon can be linear, branched or cyclic and can contain any desired number of unsaturation points. Additionally, the unsaturated hydrocarbon can exhibit conjugation. In some embodiments, the rotationally restricted linkage establishes conjugation between the two multicyclic ring moieties. One or both of the multicyclic moieties may comprise one or more heteroatoms, such as nitrogen, sulfur and/or oxygen.
Photoacoustic compounds described herein exhibit an absorption spectrum, emission spectrum and/or photoacoustic spectrum that is pH dependent. In some embodiments, absorption and emission maxima blueshift in response to decreasing pH value. Alternatively, absorption and emission maxima can redshift in response to decreasing pH value. Redshift or blueshift of absorption and emission maxima in response to increasing or decreasing pH values can be dependent on the specific compositional and structural parameters of the photoacoustic compounds. Photoacoustic compounds can be highly sensitive to changes in pH. In some embodiments, the absorption, emission and/or photoacoustic spectra of a photoacoustic compound can change in response to a change in pH value 0.1 or greater. For example, the absorption, emission and/or photoacoustic spectra of a photoacoustic compound can change in response a change in pH value of 0.1 to 0.3 or 0.1 to 0.2. Variation of photoacoustic compound spectra can result from protonation and/or deprotonation of the photoacoustic compound in response to pH change.
Additionally, absorption, emission and/or photoacoustic spectra of photoacoustic compounds can be non-responsive to other species including, but not limited to, ions of one or more alkali metals, alkaline earth metals and/or transition metals. For example, one or more spectra of a photoacoustic compound can be non-responsive to salts of Na+, K+, Ca2+, Fe3+, Fe2+, Mg2+ and/or Zn2+. In being non-responsive to various metal ionic species, photoacoustic compounds can be employed in a variety of biological and physiological environments for imaging applications based on local pH changes in the environment. Moreover, in some embodiments, photoacoustic compounds described herein are not cytotoxic, thereby further enhancing suitability of biological and physiological environments.
In some embodiments, a photoacoustic compound is of Formula (I):
wherein R1-R19 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, hydroxyl, halo, and amide, wherein the alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, and amide are optionally substituted with one or more substituents selected from the group consisting of (C1-C10)-alkyl, (C1-C10)-alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amide, halo, hydroxy, C(O)OR20, and C(O)R21, wherein R20 is selected from the group consisting of hydrogen, alkyl and alkenyl and R21 is selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl and NR22R23, wherein R22 and R23 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and heteroaryl; and wherein X is selected from the group consisting of CR24R25, N and O, wherein R24 and R25 are independent selected from the group consisting of hydrogen, alkyl, alkenyl, hydroxyl and halo.
In some embodiments, R1-R8 and R12-R19 are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, alkoxy, hydroxyl, and halo and wherein R9-R11 are independently selected from alkyl and alkenyl.
In some embodiments, a photoacoustic compound is of Formula (II):
wherein R1-R17 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, hydroxyl, halo, and amide, wherein the alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkyl-aryl, alkyl-heteroaryl, alkoxy, and amide are optionally substituted with one or more substituents selected from the group consisting of (C1-C10)-alkyl, (C1-C10)-alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy, amide, halo, hydroxy, C(O)OR18, and C(O)R19, wherein R18 is selected from the group consisting of hydrogen, alkyl and alkenyl and R19 is selected from the group consisting of hydrogen, alkyl, alkenyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl and NR20R21, wherein R20 and R21 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and heteroaryl; and wherein X is selected from the group consisting of CR22R23, N and O, wherein R24 and R25 are independent selected from the group consisting of hydrogen, alkyl, alkenyl, hydroxyl and halo.
In another aspect, methods of imaging are described. Briefly, a method of imaging a target comprises disposing a photoacoustic compound in an environment adjacent to the target and irradiating the photoacoustic compound with one or more wavelengths of electromagnetic radiation. Soundwaves produced by the photoacoustic compound in response to the electromagnetic radiation are detected and transformed into an image. The photoacoustic compound comprises two mutlicyclic ring moieties coupled by a rotationally restricted linkage, and exhibits at least one of a pH dependent absorption spectrum, pH dependent emission spectrum and pH dependent photoacoustic spectrum. In some embodiments, the photoacoustic compound is irradiated with multiple wavelengths of radiation resulting in multiple wavelength dependent images of the target. As described further herein, each of the wavelength dependent images can correspond to the photoacoustic compound at locations of differing pH in the environment. The wavelength dependent images can be overlaid to present a detailed image of the target.
In some embodiments, the target is biological tissue, and the environment is extracellular space in and/or around the tissue. The biological tissue for example, can comprise cancer cells and non-cancer cells. Due to differences in extracellular pH values (pHe), the cancer cells can be resolved from the non-cancer cells with the photoacoustic compound. Other biological or physiological environments may be imaged and characterized in this way. Additionally, non-biological environments may also be imaged with photoacoustic compounds described herein based on local pH variations within the environments. Notably, photoacoustic compounds employed in imaging methods can have any composition and/or properties described in Section I above, including structures of Formulas (I) and (II).
These and other embodiments are further illustrated in the follow non-limiting examples.
A photoacoustic compound of Formula (Ia) was synthesized and characterized.
Initial evaluation of compound (Ia) using UV-Vis spectroscopy indicated absorbance spectra modulation corresponding with variable pH PBS buffer solutions (
The quantum yield of compound (Ia) under acidic (pH 6.0), neutral (7.0), and basic (pH 8.0) buffered solutions were calculated by comparison with rhodamine (ΦR=0.95 in ethanol), where ΦF is the quantum yield. The results are provided in Table I.
As provided in Table I, absorption and emission maxima blueshift with decreasing pH value.
Since compound (Ia) was designed to spectrally alter based upon H+ protonation, compound (Ia) was also evaluated in the presence of de-ionized water, phosphate buffer saline, and/or other ions which are present in living organisms (Na+, K+, Ca2+, Fe2+, Mg2+, Zn2+, Fe3+ and Fe2+ as chloride salts), and other highly reactive cellular products (H2O2, glutathione (GSH), cysteine) at pH 7.4. The absorption spectra did not significantly alter in the presence of reactive cellular products or ions other than H+ associated with acidic pH (
Preliminary evaluation of compound (Ia) accumulation within mice was conducted. A matrigel plug containing surgicel mesh and fibroblast was injected subcutaneously to stimulate non-malignant fibrotic tissue two weeks prior to orthotopic tumor implantation in 3 SCID mice. Mice were anesthesized using 1.5% isoflurane and a gas mixture of 0.9 L medical air and 0.1 L 02 to detect tissue hypoxia. Because areas of tissue hypoxia will also have acidic extracellular pH, tissue hypoxia served as an additional acidic extracellular pH (pHe) control. One week post tumor implantation mice were imaged using MSOT to obtain a baseline prior to iv tail vein injection of compound (Ia) at (100 nM in PBS pH 7.4) in 100 μl total volume. Pancreatic tumor specific accumulation of compound (Ia) was detected after 2 h (
Various embodiments of the invention have been described in fulfillment of the various objects of the invention. It should be recognized that these embodiments are merely illustrative of the principles of the present invention. Numerous modifications and adaptations thereof will be readily apparent to those skilled in the art without departing from the spirit and scope of the invention.
The present application claims priority to U.S. Provisional Patent Application Ser. No. 62/715,550 filed Aug. 7, 2018, which is incorporated herein by reference in its entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2019/045258 | 8/6/2019 | WO | 00 |
| Number | Date | Country | |
|---|---|---|---|
| 62715550 | Aug 2018 | US |
